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HeLa cells were transfected for 24 h with pEGFP empty vector or TfR-GFP-WT or TfR-GFP-4KA treated with DMSO (basal) or Baf for 4 h, processed for immunoblot, and labeled for LC3, TfR, and GAPDH as loading control. Histogram shows relative LC3-II levels in basal or under Baf treatment, normalized for GAPDH. Data are mean ± SD (n= 4; graphs represent means from four independent experiments) one-way ANOVA with Dunnett’s multiple comparisons test,**p< 0.01,***p< 0.001; ns, not significant. See alsoFigure S3.

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