Fig. 1
From: EGF receptor kinase suppresses ciliogenesis through activation of USP8 deubiquitinase
USP8 knockdown induces trichoplein degradation and unscheduled ciliogenesis. a Screen flowchart. b, c Twenty-four hours after transfection with siRNA for IFT20 (b) or Cep164 (c), PRE1 cells were further transfected with USP8 siRNA (#1 or #2) and then cultured for 48 h in the presence of serum (10% FBS). The cells were analyzed by immunoblotting with indicated antibodies. Normalized intensities of trichoplein/GAPDH are shown as mean from three independent biological replicates. Percentages of ciliated cells and cyclin A-positive cells are shown as mean ± SD from three independent biological replicates (n > 200 each). d Control or USP8 (#1 or #2) knockdown RPE1 cells were treated with 200 nM cycloheximide-supplemented normal medium (10% FBS) for indicated times. Normalized intensities of trichoplein/GAPDH (mean ± SD from three independent technical replicates) were evaluated by immunoblotting analysis. To accurately evaluate the kinetics of trichoplein turnover, the samples were re-evaluated in Supplementary Fig. 4a. **p < 0.01, *0.01 < p < 0.05, n.s. not significant, two-tailed unpaired student’s t-tests
