- Picture from Ding WY et al. (2017) J Cell Biol "Plastin increases cortical connectivity to facilitate robust polarization and ...."
Figure S2. Expression pattern of endogenous PLST-1::GFP during embryogenesis and in the adult hermaphrodite. (A) Expression pattern of endogenousPLST-1::GFP during embryogenesis with prominent enrichment at cell-cell contacts (red arrowheads), the buccal region of the epidermis (cyan arrowhead),pharynx (green arrowhead), and intestine (magenta arrowhead). (B) In adults, PLST-1 expression was found in the socket cells of amphids (i), pharyngealintestinal valve (ii), intestinal lumen (iii), syncytial and cellularized gonad (iv), vulval cells (v), intestinal-rectal valve (vi), and socket cells of phasmids (vii).Bars: (micrograph showing whole worm) 100 um; (all others) 5 um.
- Picture from Ding WY et al. (2017) J Cell Biol "Plastin increases cortical connectivity to facilitate robust polarization and ...."
Figure 1. Endogenous PLST-1 labeled with GFP localizes to the cortex of newly fertilized C. elegans zygotes. (A) Cortical view of the newly fertilizedC. elegans zygote coexpressing PLST-1::GFP and the F-actin reporter Lifeact::RFP. Punctate and filamentous structures are highlighted by yellow and redarrowheads, respectively. Colocalization of the PLST-1::GFP and Lifeact::RFP signals is shown in the merged channel and representative band scans. (B)Pearson's correlation between PLST-1::GFP and Lifeact::RFP at various stages in single-cell zygote (n >_ 8). (C) Filamentous and punctate structures formed byPLST-1::GFP can be separated by
arx-2 and
cyk-1 RNAi. (D) Ratiometric analysis of PLST-1::GFP/Lifeact::RFP fluorescent intensities reveals that PLST-1::GFPsignal is enriched in contractile structures. (E) Time lapse of PLST-1::GFP/Lifeact::RFP during the formation and the subsequent disassembly of a contractileF-actin cluster during polarity establishment. Data are represented as mean +- SEM. Bars, 5 um.
- Picture from Ding L et al. (2000) Biochem J "Association of several small heat-shock proteins with reproductive tissues in ...."
Figure 1 Tissue distribution of small heat-shock proteinsIn all panels anterior is on the left and dorsal towards the top. Small heat-shock proteins were labelled with rabbit polyclonal antibodies as previously described [2], and the images are shown in green. (A) HSP12s (HSP12.6 antibody). A1, overview of HSP12 expression in L4 hermaphrodite. Spermathecae are strongly labelled and vulva is lightly labelled (arrows), with nuclei (red, DAPI) showing the positions of the tissues. A2-A5, staining in spermatheca (A2) is confirmed by co-staining of sperm nuclei (A3; red, DAPI). The images in A2 and A3 are merged in A4. A5, HSP12s localized in cytoplasm of sperm cells (green), with nuclei shown in red (DAPI). A6-A8, adult male showing HSP12s in sperm (S). The positions of sperm cells are indicated by nuclear counterstaining with DAPI (A7), and the distal loop of the testis (T) is indicated. Images A6 and A7 are merged in A8. A9-A12, anti-HSP12 labels vulva in adult hermaphrodites. A9 and A10, staining pattern of HSP12s in vulval region. A11, vulval muscles are revealed by counterstaining with a monoclonal antibody to myosin heavy chain, DM5.6 [16]. Images A10 and A11 are merged in A12. A13, HSP12s are ubiquitously expressed in L1 (green) with nuclei shown in red (DAPI). A14 and A15, controls: HSP12.6 antibody was pre-incubated with an excess of recombinant HSP12.3. The nematode was counterstained with DAPI to show nuclei, and with DM5.6 to label myosin. (B) HSP16 localization with polyclonal anti-HSP16-2 antibody [15]. B1, in late L1, HSP16s are ubiquitously expressed, although particularly prominent in nerve ring (arrow). B2, adult hermaphrodite: in addition to general tissue staining, anti-HSP16 strongly labels spermathecae (SP), and, more lightly, the vulval region (V). B3, HSP16s (green) are localized in sperm cytoplasm, with nuclei shown in red (DAPI). B4 and B5, HSP16s (green) are concentrated in discrete cytoplasmic regions in intestinal cells and along the border of the lumen (L); large intestinal nuclei (arrows, B5) are shown in red (DAPI). B6, HSP16s are expressed throughout the body in males, slightly concentrated over sperm (S). Sperm nuclei were stained with DAPI (B7), and the images are merged in B8. B9, unstressed L2-L3 animal, showing lack of HSP16 staining (green). The animal was counterstained with DAPI, and nuclei are shown in blue. B10, as a control for permeabilization, the animal in B9 (DAPI nuclei in blue) was also counterstained with monoclonal antibody DM5.6 [15] to show body-wall muscle (red). (C) HSP43. C1, overview of HSP43 staining pattern in male. HSP43 staining is conspicuous in the tail (T) and at hypodermal/muscle cell contacts (H), and less prominent over the location of spermatids in the testis (S). C2, anti-HSP43 labels tail rays in males (R and arrows, inset). Nuclei (red, DAPI) show the position of the tissues. C3, tail region of male, with anti-HSP43 labelling of vas deferens or, possibly, copulatory spicules (arrows). C4, HSP43 labels the hook and other structures in the male tail (arrow). C5, hermaphrodite, showing anti-HSP43 labelling of spermathecae and vulva.
- Picture from Ding L et al. (2000) Biochem J "Association of several small heat-shock proteins with reproductive tissues in ...."
Figure 1 Tissue distribution of small heat-shock proteinsIn all panels anterior is on the left and dorsal towards the top. Small heat-shock proteins were labelled with rabbit polyclonal antibodies as previously described [2], and the images are shown in green. (A) HSP12s (HSP12.6 antibody). A1, overview of HSP12 expression in L4 hermaphrodite. Spermathecae are strongly labelled and vulva is lightly labelled (arrows), with nuclei (red, DAPI) showing the positions of the tissues. A2-A5, staining in spermatheca (A2) is confirmed by co-staining of sperm nuclei (A3; red, DAPI). The images in A2 and A3 are merged in A4. A5, HSP12s localized in cytoplasm of sperm cells (green), with nuclei shown in red (DAPI). A6-A8, adult male showing HSP12s in sperm (S). The positions of sperm cells are indicated by nuclear counterstaining with DAPI (A7), and the distal loop of the testis (T) is indicated. Images A6 and A7 are merged in A8. A9-A12, anti-HSP12 labels vulva in adult hermaphrodites. A9 and A10, staining pattern of HSP12s in vulval region. A11, vulval muscles are revealed by counterstaining with a monoclonal antibody to myosin heavy chain, DM5.6 [16]. Images A10 and A11 are merged in A12. A13, HSP12s are ubiquitously expressed in L1 (green) with nuclei shown in red (DAPI). A14 and A15, controls: HSP12.6 antibody was pre-incubated with an excess of recombinant HSP12.3. The nematode was counterstained with DAPI to show nuclei, and with DM5.6 to label myosin. (B) HSP16 localization with polyclonal anti-HSP16-2 antibody [15]. B1, in late L1, HSP16s are ubiquitously expressed, although particularly prominent in nerve ring (arrow). B2, adult hermaphrodite: in addition to general tissue staining, anti-HSP16 strongly labels spermathecae (SP), and, more lightly, the vulval region (V). B3, HSP16s (green) are localized in sperm cytoplasm, with nuclei shown in red (DAPI). B4 and B5, HSP16s (green) are concentrated in discrete cytoplasmic regions in intestinal cells and along the border of the lumen (L); large intestinal nuclei (arrows, B5) are shown in red (DAPI). B6, HSP16s are expressed throughout the body in males, slightly concentrated over sperm (S). Sperm nuclei were stained with DAPI (B7), and the images are merged in B8. B9, unstressed L2-L3 animal, showing lack of HSP16 staining (green). The animal was counterstained with DAPI, and nuclei are shown in blue. B10, as a control for permeabilization, the animal in B9 (DAPI nuclei in blue) was also counterstained with monoclonal antibody DM5.6 [15] to show body-wall muscle (red). (C) HSP43. C1, overview of HSP43 staining pattern in male. HSP43 staining is conspicuous in the tail (T) and at hypodermal/muscle cell contacts (H), and less prominent over the location of spermatids in the testis (S). C2, anti-HSP43 labels tail rays in males (R and arrows, inset). Nuclei (red, DAPI) show the position of the tissues. C3, tail region of male, with anti-HSP43 labelling of vas deferens or, possibly, copulatory spicules (arrows). C4, HSP43 labels the hook and other structures in the male tail (arrow). C5, hermaphrodite, showing anti-HSP43 labelling of spermathecae and vulva.
- Picture from Ding L et al. (2000) Biochem J "Association of several small heat-shock proteins with reproductive tissues in ...."
Figure 1 Tissue distribution of small heat-shock proteinsIn all panels anterior is on the left and dorsal towards the top. Small heat-shock proteins were labelled with rabbit polyclonal antibodies as previously described [2], and the images are shown in green. (A) HSP12s (HSP12.6 antibody). A1, overview of HSP12 expression in L4 hermaphrodite. Spermathecae are strongly labelled and vulva is lightly labelled (arrows), with nuclei (red, DAPI) showing the positions of the tissues. A2-A5, staining in spermatheca (A2) is confirmed by co-staining of sperm nuclei (A3; red, DAPI). The images in A2 and A3 are merged in A4. A5, HSP12s localized in cytoplasm of sperm cells (green), with nuclei shown in red (DAPI). A6-A8, adult male showing HSP12s in sperm (S). The positions of sperm cells are indicated by nuclear counterstaining with DAPI (A7), and the distal loop of the testis (T) is indicated. Images A6 and A7 are merged in A8. A9-A12, anti-HSP12 labels vulva in adult hermaphrodites. A9 and A10, staining pattern of HSP12s in vulval region. A11, vulval muscles are revealed by counterstaining with a monoclonal antibody to myosin heavy chain, DM5.6 [16]. Images A10 and A11 are merged in A12. A13, HSP12s are ubiquitously expressed in L1 (green) with nuclei shown in red (DAPI). A14 and A15, controls: HSP12.6 antibody was pre-incubated with an excess of recombinant HSP12.3. The nematode was counterstained with DAPI to show nuclei, and with DM5.6 to label myosin. (B) HSP16 localization with polyclonal anti-HSP16-2 antibody [15]. B1, in late L1, HSP16s are ubiquitously expressed, although particularly prominent in nerve ring (arrow). B2, adult hermaphrodite: in addition to general tissue staining, anti-HSP16 strongly labels spermathecae (SP), and, more lightly, the vulval region (V). B3, HSP16s (green) are localized in sperm cytoplasm, with nuclei shown in red (DAPI). B4 and B5, HSP16s (green) are concentrated in discrete cytoplasmic regions in intestinal cells and along the border of the lumen (L); large intestinal nuclei (arrows, B5) are shown in red (DAPI). B6, HSP16s are expressed throughout the body in males, slightly concentrated over sperm (S). Sperm nuclei were stained with DAPI (B7), and the images are merged in B8. B9, unstressed L2-L3 animal, showing lack of HSP16 staining (green). The animal was counterstained with DAPI, and nuclei are shown in blue. B10, as a control for permeabilization, the animal in B9 (DAPI nuclei in blue) was also counterstained with monoclonal antibody DM5.6 [15] to show body-wall muscle (red). (C) HSP43. C1, overview of HSP43 staining pattern in male. HSP43 staining is conspicuous in the tail (T) and at hypodermal/muscle cell contacts (H), and less prominent over the location of spermatids in the testis (S). C2, anti-HSP43 labels tail rays in males (R and arrows, inset). Nuclei (red, DAPI) show the position of the tissues. C3, tail region of male, with anti-HSP43 labelling of vas deferens or, possibly, copulatory spicules (arrows). C4, HSP43 labels the hook and other structures in the male tail (arrow). C5, hermaphrodite, showing anti-HSP43 labelling of spermathecae and vulva.
- Picture from Ding L et al. (2000) J Biol Chem "HSP25, a small heat shock protein associated with dense bodies and M-lines of body ...."
Figure 6. Immunolocalization of HSP25 in C. elegans. Nematodes were stained using rabbit polyclonal antibody to HSP25 in combination with mouse monoclonal antibodies to nematode β-integrin (MH25) or to body wall muscle M-lines (MH42) (37), followed by fluorescein isothiocyanate-conjugated anti-rabbit secondary antibody and/or Texas Red-conjugated anti-mouse secondary antibody. A, anterior of an adult, stained with antibody to HSP25. Note the striated pattern seen in body wall muscle.B, localization of HSP25 to dense bodies (db) and M-lines (m); the dark line corresponds to a cell boundary (cb) with an adjacent muscle cell. C, anti-β-integrin also stains dense bodies and M-lines, as well as cell boundaries. D-F, co-localization of HSP25 and β-integrin. The staining patterns of HSP25 (D) and β-integrin (E) coincide in both the dense bodies and M-lines as shown in the superimposed image, F. G-I, co-localization of HSP25 and MH42. The staining patterns of HSP25 (G) and MH42 (H) coincide in M-lines (I). J and K, pharyngeal staining pattern of HSP25 in L2 and adult, respectively. L, staining of HSP25 at junctions between cells of the spermathecal wall.M, the animal was stained with anti-HSP25 in the presence of excess recombinant HSP25 as a competitor, together with DAPI (diamidinophenylindole) to reveal cell nuclei; note the absence of detectable HSP25 signal. N, the same animal as in panel M, with staining pattern of M-lines to ensure the animals had been permeabilized. O and P, same conditions as in M and N, respectively, at a lower magnification, showing a view of the anterior of the animal.
- Picture from Ding L et al. (2000) J Biol Chem "HSP25, a small heat shock protein associated with dense bodies and M-lines of body ...."
Figure 5. Expression of HSP25. A, effect of heat shock on HSP25 synthesis. C. elegans was cultured either at 20 C (lane 1) or heat shocked at 33 C (lane 2). Extracts were analyzed by SDS-PAGE and detected by Western blotting (upper panel, antibody to HSP25; lower panel, antibody to actin). B, extracts were prepared from C. elegans at each developmental stage and analyzed by SDS-PAGE and Western blotting. Lane 1, embryo; lanes 2-5, L1 to L4 larval stages; lane 6, adult; lane 7, recombinant HSP25. Extract loadings were normalized to actin levels (B, lower panel).
- Picture from Ding L et al. (2000) Biochem J "HSP43, a small heat-shock protein localized to specific cells of the vulva and ...."
Figure 2. Immunocytochemistry of HSP43. All animals orientated with anterior to the left unless otherwise mentioned. In all images, HSP43 staining is shown in green. (A) Young adult. HSP43 is concentrated in the vulva (v) and spermathecae (sp). (B) HSP43 expression in the adherens junctions of vulval epithelia (ventral view). HSP43 (B1) and MH27, which stains desmosomes (B2), are partially co-localized (B3). (C) Ventral view. The relative position of HSP43 in vulva (green) is compared to vulval muscle (red, arrows, phalloidin). (D) Overview of HSP43 staining in vulval region (adult, lateral view). Arrows indicate utse and 2
uv1 cells. (E1) H-shaped utse, ventro-lateral view. (E2) and (E3)
uv1 cells, with adjacent DAPI-stained gonadal nuclei (shown in red). (F) Spermathecal expression of HSP43. (F1) HSP43 staining in the spermathecal cage (green) with DAPI-stained sperm nuclei (shown in blue). (F2) HSP43 (green) is co-localized with desmosomes (red, MH27), except in spermathecal valve (arrow). (F3) More lateral view of a spermathecal cage, with HSP43 staining in green and nuclei (DAPI) in red. (F4) HSP43 staining in spermathecal valve region (green) compared with valve-muscle staining (red, phalloidin); HSP43 labels both ends of the phalloidin pattern. (G1) and (G2) L2 showing staining of muscle in red (phalloidin, G1) and nuclei in blue (DAPI, G2). The pharyngeal-intestinal valve (left arrow, G1) and intestinal-anal valve (right arrow, G1) are labelled by HSP43. (H), HSP43 staining over body-wall muscle. (H1) L3 larva. The signal is more intense where muscle cells contact one another (arrows). (H2) magnified region of L3-larval muscle surface, showing punctate pattern of HSP43 staining forming columns running circumferentially over muscle cells, probably at sites of hypodermal contacts. Columns can be resolved as doublets of elongated spots (arrows). (I) and (J) Specificity of anti-HSP43. No HSP43 signal was seen in spermatheca (I) or vulva (J) when antiserum was pre-incubated with excess pure HSP43. DAPI staining (blue) shows location of nuclei (I1 and J1), and co-staining with MH27 (red, I2) or DM5.6 (red, J2) indicates efficient permeabilization of the animals.