Figure 2: Expression pattern of RSD-3. | Scientific Reports

Figure 2: Expression pattern of RSD-3.

From: Endomembrane-associated RSD-3 is important for RNAi induced by extracellular silencing RNA in both somatic and germ cells of Caenorhabditis elegans

Figure 2

(a) Schematic representation of the genomic rsd-3::gfp expression construct. The genomic region of wild type rsd-3 including 4 kb of upstream promoter sequences and the full-length rsd-3 was C-terminally fused to gfp. Black boxes indicate coding exons of rsd-3. Gray box indicates gfp sequence. (b–g) RSD-3::GFP is ubiquitously expressed. The expression pattern of genomic rsd-3::gfp was observed. Arrowheads indicate cytoplasmic puncta. (b) Pharynx, head neurons and excretory canal (arrows). (c) Head neurons (dotted bracket). (d) Coelomocyte. (e) Intestine. (f) Body wall muscle (dotted brackets). (g) Hypodermis and seam cell. Arrows indicate seam cell nuclei. Scale bars in (b,c,e–g), 20 μm and scale bar in (d), 5 μm. (h) Schematic representation of the hypodermis-specific rsd-3::ecfp expression construct. dpy-7 promoter (431 bp) was used as hypodermis-specific promoter. Black box indicates rsd-3 cDNA. Gray box indicates ecfp sequence. (i–n) RSD-3::ECFP is associated with the TGN and endosomes in hypodermal cells. ECFP-tagged RSD-3 and Venus-tagged organelle markers were coexpressed under the dpy-7 promoter and observed in late L4 stage worms. ECFP fluorescence is pseudocolored green, and Venus fluorescence is pseudocolored red. (i,j) RSD-3::ECFP does not substantially overlap with the early endosome marker 2xFYVE::Venus both near the apical (i) and basolateral (j) plasma membranes, but structures labeled by both RSD-3::ECFP and 2xFYVE::Venus are occasionally observed (arrowheads). (k) RSD-3::ECFP shows no significant overlap with the late endosome marker Venus::RAB-7, but structures labeled by both RSD-3::ECFP and Venus::RAB-7 are occasionally observed (arrowheads). (l) RSD-3::ECFP closely associates with the medial Golgi marker AMAN-2::Venus, but not substantially overlaps each other. Most RSD-3::ECFP formes somewhat ring-like structures, and surrounds the AMAN-2::Venus-positive puncta. (m,n) RSD-3::ECFP colocalizes extensively with the TGN marker, Venus::SYN-16 (m) and Venus::CHC-1 (n). Enlarged images of the boxed areas are shown in the inset (l–n). Scale bars, 10 μm.

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