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. 2020 Oct 13;219(12):e202001031. doi: 10.1083/jcb.202001031

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© 2020 Endicott et al.

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Figure 5. — Inhibiting p110, but not Vps34, causes a reduction in the protein abundance of CMA substrates in NIH3T3 cells. NIH3T3 cells were treated for 24 h with the indicated drug doses or solvent controls. Representative Western blots with quantifications are shown for pictilisib (A), copanlisib (B), or autophinib (C). In C, SQSTM1 is shown as a positive control. Histone H3 and ACTB were used as loading controls for all experiments. Protein abundances were normalized to the geometric mean of H3 and ACTB, and then adjusted for plotting with the mean of the solvent control set to 1. n = 8 for all experiments and treatment groups. P values above brackets are derived from unpaired t tests with Welch’s correction for unequal SD. Error bars are SEM. ACTB, β-Actin; Aph, autophinib; Cop, copanlisib; H3, histone 3; Pic, pictilisib; TFA, trifluoroacetic acid.