Extended Data Fig. 1: Characterization of INTS12 phosphorylation.

a, Lysates from HeLa cells that were either untreated or treated with UV (20 J/m2, 20 min recovery), 4-NQO (5 µM, 1 h), MNNG (100 µM, 1 h), MMS (10 mM, 30 min), Anisomycin (20 µM, 1 h), Cycloheximide (100 µg/mL, 1 h) or b, HU (4 mM, 4 h), Bleomycine (10 ug/mL, 1 h), IR (10 Gy, 30 min recovery), Calicheamicin (100 nM, 1 h) as shown in b were separated on Phos-tag SDS-PAGE and immunoblotted with the indicated antibodies. c, HeLa cells expressing the indicated Integrator complex subunit were left untreated or treated with 4-NQO (5 µM, 1 h) and then analyzed as in a. d, Lysates from HeLa cells that were either untreated or treated with 4-NQO (1 µM, 1 h) and recover for the indicated time were analyzed as in a. e, Control or ZAK KO cells were untreated or treated with UV (20 J/m2, 20 min recovery) and then analyzed as in a. f, HeLa cells expressing the indicated Flag-tagged FL or truncated INTS12 proteins were untreated or treated with 4-NQO and then analyzed as in a. g, Mass spectrum of representative INTS12 peptide containing the indicated phosphorylated residue, with both b-ions and y-ions shown in green and yellow, respectively. h, HeLa cells expressing the indicated Flag-tagged mutant INTS12 proteins were untreated or treated with UV and then analyzed as in a. i, Sequence alignments of various INTS12 orthologues in different vertebrates. Green color, Serine (S)/Threonine (T) residues. Conserved phosphorylation residues are highlighted by the red boxes. j, INTS12 protein levels in control, INTS12 KO, and INTS12 KO cells reconstituted with stably expressed WT or 3 A INTS12-Flag were analyzed by WB. k, HeLa cells were depleted of INTS12 (INTS12 KO) and reconstituted with stably expressed WT or 3 A INTS12-Flag. Lysates from cells that were either untreated or treated with 4-NQO were analyzed as in a. All Western blots are representative of three independent experiments.

Source data