Extended Data Fig. 6: ERK regulates the protein stability of ETV4.
From: ETV4 is a mechanical transducer linking cell crowding dynamics to lineage specification
a, Validation of KTRs treated with specific inhibitors. n=13 cells for ERK-KTR and JNK-KTR, n=8 cells for p38-KTR pooled from two independent experiments. b,c, Representative time-course images from three independent colony-tracking assays for p38-KTR (b) and JNK-KTR (c) in H9 colonies. d, Immunofluorescence assay for pERK in H9 cells treated with PD0325901 (1 μM, 1h). n=30 cells. e, Immunofluorescence assay for pERK in large H9 colonies. Mitotic activation of ERK was seen in brightly stained cells. n=40 cells. f, ERK activity measured by the ratio of the cytoplasmic over nuclear intensities (C/N ratio) in single H9 hESCs expressing ERK-KTR under low and high density cultures (24h after seeding). n=26 cells. g, qPCR analysis for ETV4 in H9 hESCs treated with PD0325901 (1 μM). n=3 samples pooled from three independent experiments. h, Immunofluorescence assay for ETV4 in H9 hESCs treated with U0126 (10 μM, 1h). n=70 cells pooled from two independent experiments. n = number of cells (d,e,f) pooled from three independent experiments. Two-sided Student’s t-test, ***P < 0.001, **P < 0.01, *P < 0.05. Exact P values are presented in Supplementary Table 9. Scale bars: 10 µm (a), 25 µm (b,c,d,f, and h), 100 µm (e). Numerical source data and additional microscope images are available in Source data.
