Skip to main content
. 2020 Dec 18;3:789. doi: 10.1038/s42003-020-01510-2

Fig. 4. β-arrestin-1 regulates agonist-induced internal CXCR4 PTM.

Fig. 4

a Representative western blot illustrating the effect of β-arrestin-1 and 2 knockdown on CXCR4 PTM. Relative shRNA knockdown efficiency is shown in Supplementary Fig. 4a–c. b Representative western blot illustrating the effect of β-arrestin-1 knockdown on CXCL12-dependent AKT S473 phosphorylation. c Western blot quantification of CXCL12-dependent AKT S473 phosphorylation upon β-arrestin-1 knockdown. Data were normalized to phospho-AKT:total AKT and to 5 min normalized control shRNA sample. d Representative western blot illustrating the effect of β-arrestin-1 knockdown on CXCL12-dependent ERK1/2 phosphorylation. e Western blot quantification of CXCL12-dependent ERK1/2 phosphorylation upon β-arrestin-1 knockdown. Data were normalized to phospho-ERK1/2:total ERK1/2 and to 5 min normalized control shRNA sample. f, g Representative western blots illustrating total, plasma membrane, and internal pools of CXCR4 PTM upon either scramble or β-arrestin-1 shRNA knockdown. h Quantification of CXCR4 PTM at plasma membrane and internal locations upon β-arrestin-1 knockdown. CXCR4 PTMs were calculated by dividing UMB2 detection (non-posttranslationally modified CXCR4) by MYC intensity (total CXCR4) and normalized to the 0 min time point at each location. For all experiments, a minimum of three independent replicates were performed. All experiments were conducted in RPE cells overexpressing WT CXCR4 and stimulated with 12.5 nM CXCL12 for the stated time course. β-arrestin-1 knockdown experiments were conducted using two validated shRNAs (Supplementary Fig. 4). Individual data points from each experiment are plotted; mean, SD, and median line. Statistical significance *p < 0.05. Complete raw blots are shown in Supplementary Fig. 10.