Extended Data Fig. 8: SMART1-induced selective degradation of Smurf1 in vivo.
From: Targeting Smurf1 to block PDK1–Akt signaling in KRAS-mutated colorectal cancer
a, Dose–response curves for control (shCtrl) and Smurf1 silenced (shSmurf1) HCT116 cells treated with SMART1 for 72 h (n = 4 independent experiments). b, IB analysis of Smurf1 expression in HCT116 cells with Smurf1 knockdown and protein levels were quantified by ImageJ 1.8.0.345. c, Compartment of IC50 between A01, A17, SMART1Et and SMART1 (72 h) in HCT116 cells (n = 4 independent experiments). d, Representative images of SPR binding assay of SMART1Et (left) or Smurf1-L (right) to full-length Smurf1. e, SMART1 inhibited Smurf1 protein level and PDK1 neddylation in cells (Smurf1-L: 0.5 μM, SMART1Et: 0.5 μM or SMART1: 0.5 μM). IB and Nedd8 IP products and WCL derived from HEK293 cells treated with different compounds. The indicated protein levels were quantified by ImageJ 1.8.0.345. f, Effect of SMART1Et (0.5 μM, 12 h) on the proteome of HCT116 cells. Data plotted log2 of the fold change versus DMSO control against −log10 of the P-value per protein (3 independent experiments). g, MS result showed SMART1Et inhibited Akt signaling pathway. GSEA enrichment plot was displayed. h, Dose–response curves for control (sgCtrl) and CRBN KO (sgCRBN) HCT116 cells treated with SMART1 or SMART1Et for 72 h (4 independent experiments). i, Dose–response curves for control (shCtrl) and Smurf1 knockdown (shSmurf1-1/shSmurf1-2) HCT116 cells treated with SMART1 or SMART1Et for 72 h (4 independent experiments). j, IB analysis of indicated proteins in HCT116 cells with varying degrees of Smurf1 knockdown and Smurf1 protein levels were quantified by ImageJ 1.8.0.345. Immunoblots are representative of three independent experiments. Error bars denote the mean ± s.e.m.
