Fig. 2: Depletion of Arl4A accelerates EGFR mutant degradation.
From: Endosomal Arl4A attenuates EGFR degradation by binding to the ESCRT-II component VPS36
a A549, H1975 and PE089 cells were treated with either the control or Arl4A siRNA. Cell lysates were analyzed by immunoblotting with anti-EGFR and anti-α-tubulin antibodies. Arl4A knockdown efficiency was assessed by RT-PCR amplification using Arl4A-specific primers, and GAPDH was used as an internal control (lower panel). The amounts of EGFR were determined by ImageJ software. The results represent the mean ± SD of three independent experiments and the p-values were assessed by a two-sided t-test. b–d The indicated cells were transfected for 48 h with siControl or siArl4A. Cells were sequentially subjected to EGFR degradation assays as described in the “Methods” section. To compare the EGFR degradation rates, we adjusted the EGFR protein levels at time 0 in each group to be the same for western blot analysis. The relative amount of EGFR was quantified by densitometric quantification from three experiments using α-tubulin intensity to normalize the EGFR signal, and error bars represent the mean ± SD. The t1/2 ± SD of EGFR was obtained using a nonlinear one-phase decay fit of the time course; the p-values were assessed by a two-sided t-test. Source data are provided as a Source Data file.
