Figure 3. Loss of VPS13B and FAM177A1... | Rockefeller University Press

Figure 3.

Loss of VPS13B and FAM177A1 leads to delay in Golgi complex reformation after BFA treatment. (A) Anti-GM130 immunofluorescence of WT, VPS13BKO1, VPS13BKO2, FAM177A1KO, and FAM177A1; VPS13B DKO cells before incubation with BFA, after 1 h in BFA (5 µg/ml), and after subsequent washings as indicated; superscripts indicate different clones. Scale bar = 10 μm. (B and C) Western blots of total cell homogenates showing loss of the VPS13B band and/or the FAM177A1 band in the KO clones. GAPDH was used as a gel loading control. (D) Quantification of Golgi complex reformation in cells with the indicated genotypes after BFA washout for 2 or 5 h. “Rescue” refers to the exogenous expression of the knocked-out protein. Data are mean ± SEM n = 3 per condition, in each condition 75–100 cells were quantified. Unpaired, two-tailed t tests. NS, not significant. ****P < 0.0001; ***P < 0.001; **P < 0.01; *P < 0.05. Source data are available for this figure: SourceData F3.

Loss of VPS13B and FAM177A1 leads to delay in Golgi complex reformation after BFA treatment. (A) Anti-GM130 immunofluorescence of WT, VPS13BKO¹, VPS13BKO², FAM177A1KO, and FAM177A1; VPS13B DKO cells before incubation with BFA, after 1 h in BFA (5 µg/ml), and after subsequent washings as indicated; superscripts indicate different clones. Scale bar = 10 μm. (B and C) Western blots of total cell homogenates showing loss of the VPS13B band and/or the FAM177A1 band in the KO clones. GAPDH was used as a gel loading control. (D) Quantification of Golgi complex reformation in cells with the indicated genotypes after BFA washout for 2 or 5 h. “Rescue” refers to the exogenous expression of the knocked-out protein. Data are mean ± SEM n = 3 per condition, in each condition 75–100 cells were quantified. Unpaired, two-tailed t tests. NS, not significant. ****P < 0.0001; ***P < 0.001; **P < 0.01; *P < 0.05. Source data are available for this figure: SourceData F3.