FIG 5.

Characterization of ATL KO HeLa cells. (A) HeLa cells were knocked out for ATL2 and ATL3 genes (dKO) using CRISPR. HeLa WT or dKO cells were then transduced to express exogenous ATL3-myc, ATL3 WT, or ATL3 mutated in the GTPase activity domain (ATL3KA). ATL3 expression was assessed by Western blotting. Actin was used as a loading control. (B) HeLa WT or dKO cells transduced with control (ctrl), ATL3 WT, or ATL3KA vectors and expressing an ER-tracker dsRed2 were examined by confocal microscopy to observe the structure of the peripheral ER tubular network. Three-way junctions were quantified as previously described (42). At least three cells per condition per experiment from three independent experiments were analyzed and plotted. Statistical significance was determined by using a one-way ANOVA test and a Dunnett posttest. ***, P < 0.001.