Fig. 4: KCTD10 facilitates CUL3 recruitment to TRCs.
From: KCTD10 is a sensor for co-directional transcription–replication conflicts
a, Western blot of fractions from size-exclusion chromatography from control (DMSO) and ICRF193-treated (1 μM, 4 h) U2OS cells. b, KCTD10 co-immunoprecipitation assay from the indicated fractions from ICRF193-treated cells as indicated in a. c,d, PLA for CUL3–POLR2A in KCTD10-deficient U2OS cells treated with 100 nM ICRF193 or DMSO for 4 h. c, Quantification of foci. Data are mean ± s.d. (n = 200 cells examined over 3 independent experiments). d, Representative images. e,f, PLA for CUL3–POLR2A in HEK293T cells expressing the co-directional ECFP reporter treated with 1 mg ml−1 doxycycline for 20 h and 100 nM ICRF193 or DMSO for 4 h. Scale bars, 10 μm. e, Quantification of foci. Data are mean ± s.d. (n = 200 cells examined over 3 independent experiments). f, Representative images. Scale bars, 10 μm. g, iPTM scores from AlphaFold 2-multimer predictions with KCTD10. h, PAE plots for the top-ranked AlphaFold 2-multimer predictions for POLR2C–KCTD10 and TCEA2–KCTD10. Right, schematic for the uncharacterized supersecondary structure located in KCTD10 as depicted. i, Predicted structure of KCTD10 bound to POLR2C. j, Predicted structure of KCTD10 bound to TCEA2. k, Co-immunoprecipitation assay in HEK293T cells expressing Flag-tagged KCTD10 and treated with 1 μM ICRF193 or DMSO. P values were calculated with one-way ANOVA with Bonferroni’s multiple comparisons test.
