The accumulation of the Golgi membrane at the wound site is required for membrane repair. (A) Representative confocal images showing TGN-38::tagBFP before and after wounding with the treatment of rab-1 and rab-6.2 RNAi, as well as rab-6.2(ok2254) deletion mutant. Pcol-19-TGN-38::tagBFP(zjuSi280) for RNAi, and Pcol-19-TGN-38::tagBFP(zjuSi280); Pcol-19-RDE-1(juIs346); rde-1(ne219) transgenic animals were used for wounding and imaging. Scale bar: 10 µm. (B) Quantitation analysis of the intensity ratio (ΔFw/Fuw) of TGN-38::tagBFP after wounding (A). Error bars represent the mean value ± SD (n = 93, 15, 34, 58, 28 from the left to the right). One-way ANOVA multiple comparisons, ***P < 0.001. **P < 0.01. (C) Representative confocal images showing GFP::MANS-2 before and after wounding with the treatment of Blebbistatin (20 µM) and BFA (150 µg/ml). Pcol-19-GFP::MANS-2(zjuSi302) for drug treatment were used for wounding and imaging. Scale bar: 10 µm. (D) Quantitation analysis of the intensity ratio (ΔFw/Fuw) of GFP::MANS-2 after wounding (C). Error bars represent the mean value ± SD (n = 33, 45, 15, 15 from the left to the right). One-way ANOVA multiple comparisons, ***P < 0.001. (E) Quantitation analysis of the percentage of trypan blue staining of worms at 6 h after wounding treated with BFA (150 µg/ml), rab-1 RNAi, and rab-6.2 RNAi, as well as rab-6.2(ok2254) deletion mutant. N2 strain was used for drug treatments. Pcol-19-RDE-1(juIs346); rde-1(ne219) strain was used for RNAi treatment. The error bars represent the mean value. ± SD (n = 3), Student’s t test, *P < 0.05. ***P < 0.001. (F) Quantitation analysis of the survival rates (%) of animals treated with Blebbistatin (20 µM), rab-1 RNAi, rab-6.2 RNAi, and GSK-A1(100 μM) after wounding. N2 for drug treatments and Pcol-19-RDE-1(juIs346); rde-1(ne219) transgenic animals for RNAi treatments, wounding and survival rate quantification. The error bars represent the mean value ± SD (n = 3), Student’s t test, ***P < 0.001. *P < 0.05. See also Fig. S3.
