Figure S3.

Effects of PI3K inhibitors on CMA and macroautophagy in AML12 cells. (A–C) Dose curves in AML12 cells for buparlisib, pictilisib, and autophinib, respectively. (D) Western blots demonstrating the effects of buparlisib (5 µM), pictilisib (2 µM), and autophinib (5 µM) on phosphorylation of Akt, MAPKAP1, and GFAP. Blots are representative of six experimental replicates. DMSO is the solvent control for all drugs. (E and F) 10 h treatment with buparlisib or pictilisib induces the accumulation of Dendra2 CMA reporter puncta, but treatment with autophinib does not. Data were pooled from at least three independent experiments. DMSO = 202 cells, buparlisib = 122 cells, pictilisib = 104 cells, and autophinib = 95 cells. (G–I) Effects of buparlisib, pictilisib, and autophinib on macroautophagy, as measured by LC3-II flux, n = 6. For all experiments, cells were maintained in complete growth medium (see Materials and methods). P values written above brackets are derived from unpaired t tests. Two-way ANOVA tables are displayed below graphs where appropriate. Error bars are SEM. Scale bars are 20 µm. ACTB, β-Actin; Aph, autophinib; Bup, buparlisib; Pic, pictilisib.