Picture from Yoshina S et al. (2022) MicroPubl Biol "Integration of multicopy extrachromosomal transgenes into defined loci ...."

Figure 1. Isolation of multicopy integrant strains by the CRISPR/Cas9 system.(A) Procedures for isolation of multicopy integrant strains by the CRISPR/Cas9 system. Fluorescent markers are shown by green (EGFP) and red (DsRed). (B) Relative amounts of the lag-2 promoter and the T07F10.1 gene as determined by quantitative PCR. Primer set 1 is targeted at the lag-2 promoter. Primer set 2 is targeted at the T07F10.1 gene. The data were normalized with the ama-1 gene. The copy numbers are presented as a ratio to the wild-type N2 (n>3). (C) A scatter plot of the copy number of integrated sequences (abscissa) against fluorescence intensity (ordinate) in the Ex5672, and the 14 Is lines. The X axis is shown in log scale. The graph represents the intensity of fluorescence of DTC (y-axis, n > 20 animals per strain). Fluorescence intensity was measured using ImageJ (NIH, Bethesda, MD). (D) Example of GFP protein expressed in DTC. Scale bar = 5.0 μm. (2 sec. exposure time, gain X 4)

Picture from Yoshina S et al. (2017) MicroPubl Biol "Novel deletion alleles of a C. elegans gene C38D4.9, named as tm4476 and ...."

We report tm4476 and tm4561 as novel alleles of C38D4.9 gene that is an ortholog of human METTL5 (methyltransferase like 5)1. Human METTL5 function has not been reported. The alleles were isolated from the comprehensive screening of gene deletions generated by TMP/UV2. In the screening, both the alleles were detected by nested PCR using the following primer sets, 5'-CCGCCTATATCATGGCGCTT-3' and 5'-GCTGGTAGTTGCCACCGCAT-3' for first round PCR and 5'-ACTATCGAAGCCGCGCTTCA-3' and 5'-AACCCCAACTCCGTCCCATT-3' for second round PCR. By Sanger sequencing, the 30 bp flanking sequences of the alleles tm4476 and tm4561 were identified as GTATGAGTTGGAGACTGTTTTAGGAGTTGA-(191 bp deletion)-GTACTCTTTAAAAGCGCACATCTTTCTGAA and TAATTGATATCGGATGTGGATGTGGAATGT-(309 bp deletion)-ATTTTCGGCTAAAAATCGTATTTGGGTAAT, respectively (Fig. 1). Based on the information about the splicing isoforms of C38D4.9 (WormBase3 WS259), the second exon of C38D4.9 is deleted in tm4476 and tm4561.

Picture from Yoshina S et al. (2015) PLoS One "Loss of C. elegans GON-1, an ADAMTS9 Homolog, Decreases Secretion Resulting in ...."

Fig 6. Depletion of gon-1 causes DAF-7 secretion defects. (a, b) Fluorescent images of DAF-7::mCherry, and DIC images (inset) in the head neurons of a wild-type animal (a) and a gon-1(tm3146) mutant animal (b) (160 msec. exposure time). The scale bar indicates 10 um. (c) Quantification of the mCherry fluorescence in neurons. The graph represents the combined results of 3 independent experiments (n > 20 neurons per strain). *p < 0.05. Bars represent themean SE. (d, e) Representative fluorescent images of DAF-7::mCherry in the coelomocytes of wild-type (d) and a gon-1(tm3146) mutant (e) animals (100 msec. exposure time). Coelomocytes are outlined with dotted circles in the fluorescent images. Insets show DIC images. The scale bar indicates 10 um. (f) Quantification of the mCherry fluorescence in coelomocytes. The graph represents the combined results of 3 independent experiments (n > 30 coelomocytesper strain). Fluorescence intensity was examined using ImageJ (NIH, Bethesda, MD). *p < 0.05. Bars represent the mean SE.IL is WBbt:0003888" Certain

Picture from Yoshina S et al. (2015) PLoS One "Loss of C. elegans GON-1, an ADAMTS9 Homolog, Decreases Secretion Resulting in ...."

Expression patterns of gon-1p::GFP.(a, c, e, g), GFP fluorescence; (b, d, f, h), DIC images. gon-1 is expressed in motorneurons at the adult stage (a), intestine at the L4 stage (c), vulval muscle, VC4 and VC5at the adult stage (e). The white arrow indicates VC4. The white arrowhead indicatesVC5. The asterisk indicates vulval muscle. gon-1 is also expressed in some cells at theembryonic stage (260-280 min after the first cleavage) (g). The scale bars indicate 20 um.

Picture from Yoshina S et al. (2015) PLoS One "Loss of C. elegans GON-1, an ADAMTS9 Homolog, Decreases Secretion Resulting in ...."

Fig 1. gon-1 is expressed in a subset of neurons. (a, c), GFP fluorescence; (b, d), DIC image. (a) GFP is observed in some head neurons (arrowheads), intestine, and excretory cells in the larva. (c) GFP is observed in a CAN neuron (arrowhead), an excretory canal cell, and body wall muscle cells.

Picture from Yoshina S et al. (2015) PLoS One "Loss of C. elegans GON-1, an ADAMTS9 Homolog, Decreases Secretion Resulting in ...."

(e, f) Representative fluorescent images of gon-1p::mCherry (e) and mgIs40[daf-28p::GFP] (f). (g) The merged image. (h) The DIC image of the same fieldshown in the (e-g). The scale bar indicates 20 1/4m. The white arrowhead indicates an ASI neuron. The white arrow indicates an ASJ neuron. The pinkarrowhead indicates an unidentified neuron.

Picture from Yoshina S et al. (2015) PLoS One "Loss of C. elegans GON-1, an ADAMTS9 Homolog, Decreases Secretion Resulting in ...."

Fig 5. GON-1 and DAF-28 are expressed in the same cells. (a, b) Representative fluorescent images of a gon-1(tm3146) mutant animal co-expressinggon-1p::GFP (a) and daf-28p::daf-28::mCherry (b). (c) The merged image. (d) The DIC image of the same field shown in (a-c). The scale bar indicates 5 1/4m.(e, f) Representative fluorescent images of gon-1p::mCherry (e) and mgIs40[daf-28p::GFP] (f). (g) The merged image. (h) The DIC image of the same fieldshown in the (e-g). The scale bar indicates 20 1/4m. The white arrowhead indicates an ASI neuron. The white arrow indicates an ASJ neuron. The pinkarrowhead indicates an unidentified neuron.

Picture from Meng L et al. (2016) PLoS Genet "Regulation of Gap Junction Dynamics by UNC-44/ankyrin and UNC-33/CRMP through ...."

Representative images of immunostaining results for FLAG::UNC-33(S)(A) and UNC-44(B) in control and mutant animals. UNC-33(S) localization was determined by a transgene expressing FLAG tagged UNC-33(S) in all neurons (Prgef-1::FALG::unc-33(S)).

Picture from Meng L et al. (2016) PLoS Genet "Regulation of Gap Junction Dynamics by UNC-44/ankyrin and UNC-33/CRMP through ...."

Representative images of immunostaining results for FLAG::UNC-33(S)(A) and UNC-44(B) in control and mutant animals. UNC-33(S) localization was determined by a transgene expressing FLAG tagged UNC-33(S) in all neurons (Prgef-1::FALG::unc-33(S)).

Picture from Le Bot N et al. (2003) Curr Biol "TAC-1, a regulator of microtubule length in the C. elegans embryo."

(B) Still images from a time-lapse movie of an embryo expressing GFP:TAC-1. The pictures show GFP:TAC-1 behavior from prophase in the 1-celled embryo (just after pronuclear-centrosome rotation) to the 2-cell stage. (0 s) prophase, (+85 s) metaphase, (+130 s) anaphase, (+140 s) telophase, (+700 s) early 2-celled embryo, (+805 s) interphase 2-celled embryo. The scale bar represents 10 um.(C) GFP:TAC-1 around the chromosomes on a metaphase plate of a multicellular embryo. The scale bar represents 5 um.