Fig. 6: FDR-1 interacts with DPY-11, critical for maintaining UPRER. | Nature Communications

Fig. 6: FDR-1 interacts with DPY-11, critical for maintaining UPRER.

From: The shutdown of food digestion due to endoplasmic reticulum homeostasis disruption acts as a protective mechanism in C. elegans

Fig. 6

a Immunoprecipitation of FDR-1::GFP::FLAG from control (N2) and fdr-1p::fdr-1::gfp::flag transgenic animals. b Microscope images showing co-localization of FDR-1 (GFP) with DPY-11 (mCherry) in the C. elegans gut. Scale bar, 10 µm. c In vivo immunoprecipitation (IP) experiment validating the interaction between FDR-1 and DPY-11 using transgenic animals expressing fdr-1p::fdr-1::gfp::flag (#1) and fdr-1p::fdr-1::gfp::flag + dpy-11p::dpy-11::myc::mCherry (#2). Transgenic animals (#1) were used as a negative control. For co-immunoprecipitation, lysates from transgenic worms (#1 and #2) were incubated with anti-FLAG beads to pull down FDR-1::GFP::FLAG binding proteins. The binding proteins were then eluted and detected by Western blot (anti-myc). The top panel shows FDR-1::GFP::FLAG and DPY-11::myc::mCherry. Adult animals cultured under standard E. coli OP50 conditions were collected for these experiments. d Representative images and quantification of GFP expression in the CPL-1(W32A Y35A)::YFP reporter strain at day 1 of adulthood, treated with control or dpy-11 RNAi. Scale bar, 100 μm. Mean ± SD; ****p < 0.0001 via two-sided t-test. e Representative images and quantification of GFP expression in wild-type animals carrying the hsp-4p::gfp reporter at the young adult stage, treated with control RNAi or dpy-11 RNAi. Scale bar, 100 μm. Mean ± SD; ****p < 0.0001 via two-sided t-test. f Developmental progression of N2, dpy-11(CB207)and dpy-11(CB1180) animals grown on heat-killed E. coli + S. saprophyticus at 20 °C for 4 days. Scale bar, 200 μm. Mean ± SD; ****p < 0.0001 via two-sided t-test. g, h Microscope images (g) and Western blot analysis (h) showing GFP expression in the fdr-1p::fdr-1::gfp reporter strain at the young adult stage, treated with control RNAi or dpy-11 RNAi. Scale bar, 100 μm (g). The relative intensity of FDR-1::GFP::FLAG versus tubulin is quantified and shown in (h). Mean ± SD; ** P < 0.01, ****p < 0.0001 via two-sided t-test. For all panels, n = number of animals which were scored. Representative data shown is one of three biological replicates. Source data are provided as a Source Data file. See also Supplementary Fig. 8 and Supplementary Data 3.

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