FIGURE 3.

TPC2 overexpression did not affect the early autophagic progression in HeLa cells. A, bafilomycin (10 nm, 1 h) only induced the accumulation of LC3-II and p62 in control, not TPC2-overexpressing, HeLa cells, whereas starvation induced autophagic progression in both cells. The quantification of the four boxed lanes is shown. B, starvation (60 min) induced RFP-ATG5 puncta, most of which were not colocalized with GFP-LC3 puncta, in both control and TPC2-overexpressing HeLa cells. C, TPC2 failed to induce LC3-II and p62 in ATG5 knockdown cells. D, ATG5 knockdown markedly inhibited TPC2-induced accumulation of LC3-II and p62 in HeLa cells. E, TPC2 overexpression did not affect p62 incorporation into autophagosome in HeLa cells. Scale bar = 10 μm.