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. 2022 Sep 28;221(11):e202110132. doi: 10.1083/jcb.202110132

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© 2022 Jani et al.

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Figure 5. — Depletion of PI4KIIs prevents the initiation and elongation of early endosomal tubules. (A and B) HeLa cells treated with control (A) or PI4KIIs siRNAs (B) were immobilized by high-pressure freezing, and ultrathin sections were analyzed by conventional EM. Arrowheads point to nascent tubular structures emerging from the limiting membranes of endosomes. Arrows (B) point to endosomes without vesicular or tubular budded structure. Insets are additional representative endosomal structures captured in cells as in A and B. (C) Quantification of the average number of nascent tubules per endosome in siCTRL- and siPIK4IIs-treated HeLa cells as in A and B. (D) Quantification of the average maximum length of endosomal nascent tubules in siCTRL- (n = 40) and siPI4KIIs- (n = 47) treated HeLa cells measured in C. Data are the average of three independent experiments presented as the mean ± SEM (siCTRL, n = 8 cells; siPI4KIIs, n = 14 cells). (C and D) Two-tailed unpaired t test; *,P < 0.05; ****, P < 0.0001. Scale bars: (main panels) 1 μm; (insets) 200 nm.