Figure S5.
Supplementary material toFigs. 5, 6, and 7. (A and B) TauRD (P301L)-Venus aggregate–positive cells stably expressing mCherry, mCherry-tagged sgRNA-resistant PTPN23 proteins illustrated in Fig. 5 A (A), or mCherry-tagged sgRNA-resistant UBAP1 proteins illustrated in Fig. 6 A (B) were transfected with control or UBAP1 sgRNA (A), or control or PTPN23 sgRNA (B), and successfully transfected cells were selected by puromycin selection. Whole-cell lysates were immunoblotted with the indicated antibodies 1 wk after transfection. (C) Representative image of α-Syn (A53T)-Venus aggregate–positive cells. Scale bar: 10 μm. (D and E) α-Syn (A53T)-Venus aggregate–positive cells were transfected with control, TSG101, CHMP4A/B/C, or VPS4A/B siRNA (D), or transfected with control or PTPN23 sgRNA, and successfully transfected cells were selected by puromycin treatment (E). α-Syn (A53T)-Venus fluorescence was measured by flow cytometry 72 h (D) or 1 wk (E) after transfection. Data were normalized to the Venus fluorescence of siControl (D)- or sgControl (E)-treated cells and represent the mean ± SEM (n = 3, from three independent experiments). Significance was calculated using one-way ANOVA with Dunnett’s test (D) or two-tailed Student’s t test (E) with *P < 0.05, **P < 0.01, ***P < 0.001. (F) Representative image of PolyQ97-Venus aggregate–positive cells. Scale bar: 10 μm. (G and H) PolyQ97-Venus aggregate–positive cells were transfected with control, TSG101, CHMP4A/B/C, or VPS4A/B siRNA (G), or transfected with control or PTPN23 sgRNA, and successfully transfected cells were selected by puromycin treatment (H). PolyQ97-Venus fluorescence was measured by flow cytometry 72 h (G) or 1 wk (H) after transfection. Data were normalized to the Venus fluorescence of siControl (G)- or sgControl (H)-treated cells and represent the mean ± SEM (n = 3, from three independent experiments). Significance was calculated using one-way ANOVA with Dunnett’s test (G) or two-tailed Student’s t test (H) with **P < 0.01, ***P < 0.001. n.s., not significant. Source data are available for this figure: SourceData FS5.

Supplementary material to Figs. 5, 6, and 7. (A and B) TauRD (P301L)-Venus aggregate–positive cells stably expressing mCherry, mCherry-tagged sgRNA-resistant PTPN23 proteins illustrated in Fig. 5 A (A), or mCherry-tagged sgRNA-resistant UBAP1 proteins illustrated in Fig. 6 A (B) were transfected with control or UBAP1 sgRNA (A), or control or PTPN23 sgRNA (B), and successfully transfected cells were selected by puromycin selection. Whole-cell lysates were immunoblotted with the indicated antibodies 1 wk after transfection. (C) Representative image of α-Syn (A53T)-Venus aggregate–positive cells. Scale bar: 10 μm. (D and E) α-Syn (A53T)-Venus aggregate–positive cells were transfected with control, TSG101, CHMP4A/B/C, or VPS4A/B siRNA (D), or transfected with control or PTPN23 sgRNA, and successfully transfected cells were selected by puromycin treatment (E). α-Syn (A53T)-Venus fluorescence was measured by flow cytometry 72 h (D) or 1 wk (E) after transfection. Data were normalized to the Venus fluorescence of siControl (D)- or sgControl (E)-treated cells and represent the mean ± SEM (n = 3, from three independent experiments). Significance was calculated using one-way ANOVA with Dunnett’s test (D) or two-tailed Student’s t test (E) with *P < 0.05, **P < 0.01, ***P < 0.001. (F) Representative image of PolyQ97-Venus aggregate–positive cells. Scale bar: 10 μm. (G and H) PolyQ97-Venus aggregate–positive cells were transfected with control, TSG101, CHMP4A/B/C, or VPS4A/B siRNA (G), or transfected with control or PTPN23 sgRNA, and successfully transfected cells were selected by puromycin treatment (H). PolyQ97-Venus fluorescence was measured by flow cytometry 72 h (G) or 1 wk (H) after transfection. Data were normalized to the Venus fluorescence of siControl (G)- or sgControl (H)-treated cells and represent the mean ± SEM (n = 3, from three independent experiments). Significance was calculated using one-way ANOVA with Dunnett’s test (G) or two-tailed Student’s t test (H) with **P < 0.01, ***P < 0.001. n.s., not significant. Source data are available for this figure: SourceData FS5.

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