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. 2021 Mar 4;2021:10.17912/micropub.biology.000373. doi: 10.17912/micropub.biology.000373

Figure 1.

Figure 1

A. Patient mutation and alignment of region of interest in human COG4 and C. elegans COGC-4. B. No distal tip cell migration defects observed in adult animals. The distal tip cell marker used allows visualization of the plasma membrane of lag-2 expressing cells, specifically of the distal tip cell. C. No co-localization of ER (SP12::GFP) and Golgi (ugtp-1::mChr) markers observed in 2-cell embryos. D. Paraquat Survival Assay. Time points represent hours after young L4 worms were placed in a 96-well plate containing various concentrations of Paraquat in M9. Each well was stirred for about 3 seconds with a wooden toothpick before worms were counted as either dead or alive. Worm counts were done at the beginning (immediately following the ‘0 hour’ counts) and after 8 hours. Worms were counted as “alive” if visibly thrashing, or if any type of movement was observed following “stirring” or directly after being poked with the wooden stick. Similarly, worms were counted as “dead” if no movement at all was observed for 5-10 seconds after stirring the well or poked directly. Sample size was 5-10 worms per well, 4 replicates per condition. Survival was compared at each Paraquat concentration using two-way ANOVA (genotype x timepoint) with Sidak multiple comparison’s test. There was no significant difference in survival between N2 and cogc-4 worms. E. Heat shock: Water Bath. Percentage of N2 and cogc-4(av107[G529R]) L4 stage worms that were alive and visibly crawling 24 and 96 hours, respectively, after 3 hours at three different heat shock temperatures (37°C, 38.5°C and 40°C). Sample size was 30-50 worms per plate, 3 replicates per temperature. Visible thrashing after each heat shock was compared using two-way ANOVA (genotype x temperature) with Sidak multiple comparison’s test. There were significantly less N2 worms visibly thrashing following the 3 hour water bath at 38.5°C, only (*P = 0.0078). F. Heat Shock: Temperature Gradient. Across all eight temperatures assayed, both N2 and cogc-4(av107[G529R]) worms displayed similar levels of response immediately following heat shock (30 minute recovery) and 24 hours after the heat shock (24 hour recovery). Sample size was 5-10 worms per tube, 3 replicates per temperature. Visible thrashing was compared using two-way ANOVA (genotype x temperature) with Sidak multiple comparison’s test. There was no significant difference in survival between N2 and cogc-4 worms.