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. 2021 Apr 22;16(5):1276–1289. doi: 10.1016/j.stemcr.2021.03.030

Figure 1.

Figure 1

Generation and validation of human iPSCs and iPSC-derived cortical neurons depleted of SQSTM1

(A) Representative fluorescence images of SQSTM1 KO and unedited WT iPSCs (RBi001-A) immunostained against SQSTM1 (green) and Hoechst (blue). Scale bars, 50 μm.

(B) Representative fluorescence images of cortical neurons, after 80 days of differentiation, immunostained against SQSTM1 (green), MAP2 (red), CTIP2 (green), TUBB3/TUJ1 (red), TBR1 (green), SLC17A7/VGlut1 (green), and Hoechst (blue). Scale bars, 25 μm.

(C) Relative mRNA expression of markers for cortical brain regions and pan-neuronal genes. Dots indicate expression mean ± SEM of neurons generated from two independent neural differentiation.

(D) Representative western blot analysis of LC3-II under normal or starvation conditions, treated with or without Baf-A1 overnight. β-Actin was used as a loading control.

(E and F) Bar graphs showing the band intensity ratio of LC3-II to β-actin detected in iPSC-derived neurons cultured in full medium (E) or starvation medium (F). Data are expressed as mean ± SEM of three independent experiments. Statistical differences were tested by unpaired two-tailed t test with Welch's correction. ∗p < 0.05.