Extended Data Fig. 4: Genetic and chemical disruptions of actin induces RPAC levels.
From: Actin remodelling controls proteasome homeostasis upon stress
Representative microscopy images (maximum intensity Z-projection) of WT (ACT1) and act1-101 cells either grown at the permissive temperature (25 °C) or shifted to the non-permissive temperature (37 °C) for 4 h and stained with Rhodamine phalloidin to visualise actin (hot red LUT). Scale bars, 3 μm. n = 3 biologically independent experiments. b, Western blot analysis of RPACs in WT and act1-101 cells either grown at the permissive temperature (25 °C) or shifted to the non-permissive temperature (37 °C) for 4 h. Ponceau S staining was used as a loading control. n = 3 biologically independent experiments. c, Representative microscopy images (maximum intensity Z-projection) of WT cells treated or not with either 25 μM Latrunculin-B or 12.5 μM Latrunculin-A for 3 h and stained for actin (hot red LUT). Scale bars, 3 mm. n = 3 biologically independent experiments. d, Frequency of ADC17 mRNAs colocalising with Ede1-tdimer2 in cells treated with either 25 μM Latrunculin-B (Lat-B) or 12.5 μM Latrunculin-A (Lat-A) for 3 h. Data are presented as mean ± s.d., n = 4 biologically independent experiments (>500 ADC17 mRNAs per condition). Statistical analysis was carried out using unpaired two-tailed Student’s t-test. ns not significant. e, Western blot analysis of RPACs and Mpk1 kinase in WT cells treated or not with either 25 μM Latrunculin-B (Lat-B) or 12.5 μM Latrunculin-A (Lat-A) for 3 h. Ponceau S staining was used as a loading control. n = 3 biologically independent experiments.
