Figure S3. cima-1
Is Expressed and Required in Epidermal Cells, Related to Figure 4 (A-H), CIMA-1(genomic)::SL2::GFP is expressed in the embryo (A, B), larval (C, D), and adult stages (E-H) in the intestine (D, as indicated by the arrow) and in epidermal cells (D, E and G, as indicated by arrow heads), but not in the AIY interneuron (E, F) or VCSC glia (G, H) (as indicated by asterisk). The AIY interneuron and the VCSC glia were colabeled with cytoplasmic mCherry (F, H). (I) Cell-specific expression of cima-1
cDNA in neurons/intestine (aex-3
promoter) and in VCSC glia (hlh-17
promoter) does not result in rescue. However, endogenous expression (cima-1
promoter) or tissue-specific expression in epidermal cells (rol-6
promoter) rescues the AIY presynaptic defect. Pcol-19 promoter drives expression after larval stages and in epidermal cells (Cox and Hirsh, 1985; Liu and Ambros, 1991). Therefore rescue using the Pcol-19 suggests that cima-1
acts postdevelopmentally in epidermal cells. Error bars are SEM. (J-Y) CIMA-1 protein fused to RFP (CIMA-1::RFP) is coexpressed in epidermal cells with mitochondrial marker TOM20::YFP (J-M), early endosomal marker GFP::RAB-5 (N-Q), later endosome GFP::RAB-7 (R-U), and lysosomal marker GFP::CUP-5 (V-Y). White arrows indicate CIMA-1::RFP, arrow heads indicate GFPmarked proteins and orange arrows indicate the colocalization of CIMA-1::RFP and GFP-marked proteins. Notice that CIMA-1::RFP is largely colocalized with lysosomal marker GFP::CUP-5. Consistent with this, Pearson's correlation coefficient for CIMA-1 and CUP-5 is 0.53 (for the other markers: CIMA-1 and TOM20 is 0.021; CIMA-1 and RAB-5 is 0.22; CIMA-1 and RAB-7 is 0.28). The scale bar in J applies to N, R, and V; scale bar in K applies to L-M, O-Q, S-U and W-Y.