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. 2021 Jan 14;220(2):e202004229. doi: 10.1083/jcb.202004229

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© 2021 Kulasekaran et al.

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Figure S5. — EGFR recycling in U87 glioblastoma cells. (A) U87 glioblastoma cells were transduced with a control (ctrl) shRNAmiR or shRNAmiRs targeting Rab35. After 3 d in culture, RNA was extracted from the cells and analyzed for the levels of Rab35 using quantitative PCR. Data are shown as mean ± SEM. Statistical analysis employed an unpaired t test. ****, P < 0.0001; n = 3 (B) U87 glioblastoma cells were transduced with a control (ctrl) shRNAmiR or a shRNAmiR targeting Rab35. The cells were then treated with DMSO or 100 µg/ml cycloheximide for 16 h. Cells were processed for immunoblot with the indicated antibodies. The migration of molecular mass markers (in kD) is indicated. (C) Quantification of total EGFR (T-EGFR) from experiments as in B. Data are shown as mean ± SEM. Statistical analysis employed a two-way ANOVA followed by Bonferroni’s multiple comparisons test; n = 3; ***, P < 0.001; *, P < 0.05; ns, not significant. (D) Data from B normalized to the respective control. Data are shown as mean ± SEM. Statistical analysis employed a two-way ANOVA followed by Bonferroni’s multiple comparisons test; n = 3; ***, P < 0.001; **, P < 0.01; *, P < 0.05.