Figure 3.

SUMOylated DPCs accumulate in SPRTN-depleted cells

(A) Parental and ΔSPRTN HeLa cells were treated with 1.35 mM FA for 10 min at 37°C and allowed to recover for the indicated times. Total DPCs were isolated by RADAR and analyzed by western blot for the indicated PTMs. Graphs show the mean ± SEM of the relative signal from three independent experiments.

(B) HeLa cells were treated with 1.35 mM FA for 10 min at 37°C. No FA was added to ΔSPRTN HeLa cells. Cells were pre-extracted, fixed, and immunostained with the indicated antibodies. Foci were counted from EdU-positive cells (200 nuclei) with ImageJ, and statistical significance was calculated using and unpaired t test.

(C) Normal MRC5 (CTRL) and RJALS patient B-II:1 primary fibroblasts were pre-extracted, fixed, and immunostained with the indicated antibodies. The SUMO-1 signal was quantified in EdU-positive cells (25 nuclei) using ImageJ, and statistical significance was calculated using an unpaired t test.

See also Figure S2.