Picture from Winston WM et al. (2007) Proc Natl Acad Sci U S A "Caenorhabditis elegans SID-2 is required for environmental RNA ...."

Figure 2. SID-2 is a predicted membrane protein conserved in C. briggsae and C. remanei, and SID-2::GFP is expressed in the luminal intestinal membranes. (A) Translated sequence of C. elegans sid-2 cDNA aligned with a conceptual assembly of the homologous C. briggsae and C. remanei genes. Identical amino acids are marked with an asterisk and similar amino acids are marked with a colon or period. The predicted signal peptide and transmembrane domain are underlined. The positions and amino acid sequence changes for five sequenced alleles are shown above the sequences (asterisk, stop). (B) Localization of SID-2::GFP expression to intestinal lumen. Fluorescent, white light, and overlay respectively at low (Upper) and high (Lower) magnification. (Scale bars, 0.1 mm and 10 um.)

Picture from Jang S et al. (2016) Neuron "Glycolytic Enzymes Localize to Synapses under Energy Stress to Support Synaptic ...."

(A and A0) PFK-1.1::eGFP (pseudocolored magenta) localization in NSM after approximately 10 min (A) and 30 min (A0) of exposure to hypoxic conditions (see Supplemental Experimental Procedures). Clustering dynamics of PFK-1.1 vary between individual animals, but clustering of PFK-1.1 is observed for most animals within 10 min of being exposed to hypoxic conditions. Here we filmed a neuron with slow clustering dynamics to capture the time-lapse image. Cell body of NSM neuron is indicated by asterisk. The time-lapse images of the inset in (A) and (A0) (taken at 1 min intervals) are displayed below. See Movie S2 for time-lapse movie.(B and B0) Pixel fluorescence values for the first inset in the montage (B) (yellow trim in bottom row) and the last inset (B0) (orange trim in bottom row). (C-E) PFK-1.1::eGFP (pseudocolored magenta) and mCherry::RAB-3 (pseudocolored green) simultaneously visualized in AIY interneurons. PFK-1.1 in AIY after 20 min (C) and 40 min (C0) of hypoxic conditions, and (C0) visualized with synaptic vesicle marker RAB-3 (D); the brightness of (C) was increased more than in (C0) in order to allow for visibility of the diffuse localization of PFK-1.1. Note that PFK-1.1 becomes increasingly clustered to presynaptic sites (denoted with brackets as ''synaptic region'') and is excluded from asynaptic regions (Colon-Ramos et al., 2007; White et al., 1986). Inset in (D) corresponds to zoomed-in (53) image of PFK-1.1 cluster localizing near a pool of synaptic vesicles. Enrichment of PFK-1.1 clustering in the AIY neurite over time in hypoxic treatment was quantified(N = 12) (see Supplemental Experimental Procedures) (E). tinitial is >20 min and tfinal is 20-40 min of hypoxic treatment. Each circle represents an individual animal in the longitudinal study.(F) Quantification of the number of PFK-1.1 clusters in the synaptic region and the asynaptic regions of AIY (N = 15).(G) Pixel fluorescence values of PFK-1.1 and RAB-3 along the neurite for boxed region in (D) to compare the colocalized distribution of PFK-1.1 relative to synaptic vesicle proteins. Left y axis corresponds to PFK-1.1, and right y axis corresponds to RAB-3 pixel fluorescence values.(H) Quantification of the distance between the maximum pixel fluorescence values of PFK-1.1 and the maximum pixel fluorescence values of RAB-3 in AIYneurons (n = 26).

Picture from Jang S et al. (2016) Neuron "Glycolytic Enzymes Localize to Synapses under Energy Stress to Support Synaptic ...."

Figure 4. Glycolytic Proteins Dynamically Cluster Near Presynaptic Release Sites(A and A0) PFK-1.1::eGFP (pseudocolored magenta) localization in NSM after approximately 10 min (A) and 30 min (A0) of exposure to hypoxic conditions (see Supplemental Experimental Procedures). Clustering dynamics of PFK-1.1 vary between individual animals, but clustering of PFK-1.1 is observed for most animals within 10 min of being exposed to hypoxic conditions. Here we filmed a neuron with slow clustering dynamics to capture the time-lapse image. Cell body of NSM neuron is indicated by asterisk. The time-lapse images of the inset in (A) and (A0) (taken at 1 min intervals) are displayed below. See Movie S2 for time-lapse movie.(B and B0) Pixel fluorescence values for the first inset in the montage (B) (yellow trim in bottom row) and the last inset (B0) (orange trim in bottom row). (C-E) PFK-1.1::eGFP (pseudocolored magenta) and mCherry::RAB-3 (pseudocolored green) simultaneously visualized in AIY interneurons. PFK-1.1 in AIY after 20 min (C) and 40 min (C0) of hypoxic conditions, and (C0) visualized with synaptic vesicle marker RAB-3 (D); the brightness of (C) was increased more than in (C0) in order to allow for visibility of the diffuse localization of PFK-1.1. Note that PFK-1.1 becomes increasingly clustered to presynaptic sites (denoted with brackets as ''synaptic region'') and is excluded from asynaptic regions (Colon-Ramos et al., 2007; White et al., 1986). Inset in (D) corresponds to zoomed-in (53) image of PFK-1.1 cluster localizing near a pool of synaptic vesicles. Enrichment of PFK-1.1 clustering in the AIY neurite over time in hypoxic treatment was quantified(N = 12) (see Supplemental Experimental Procedures) (E). tinitial is >20 min and tfinal is 20-40 min of hypoxic treatment. Each circle represents an individual animal in the longitudinal study.(F) Quantification of the number of PFK-1.1 clusters in the synaptic region and the asynaptic regions of AIY (N = 15).(G) Pixel fluorescence values of PFK-1.1 and RAB-3 along the neurite for boxed region in (D) to compare the colocalized distribution of PFK-1.1 relative to synaptic vesicle proteins. Left y axis corresponds to PFK-1.1, and right y axis corresponds to RAB-3 pixel fluorescence values.(H) Quantification of the distance between the maximum pixel fluorescence values of PFK-1.1 and the maximum pixel fluorescence values of RAB-3 in AIYneurons (n = 26). (I-P) PFK-1.1::mCherry (pseudocolored magenta) (I and M) and glycolytic proteins GPD-3::eGFP (J) or ALDO-1::eGFP (N) were simultaneously visualized in NSM neurons (K and O) under hypoxic conditions. Quantification of the distance between the maximum pixel fluorescence values of PFK-1.1 and GPD-3 (L) (n = 11) and PFK-1.1 and ALDO-1 (P) (n = 23).Scale bar, 5 um. Error bars: SEM. *p < 0.05, **p < 0.01, ***p < 0.001.