Anti mpk1

Anti-Adc17 (Bertolotti laboratory; rabbit; 1:1,000), anti-Adc17-(2) (DSTT; sheep; 1:250; DU66321), anti-Nas6 (Abcam; rabbit; 1:2,000; ab91447), anti-Flag (Sigma Aldrich; mouse; 1:2,000; F3165), anti-Rpt5 (Enzo Life Sciences; rabbit; 1:5,000; PW8245), anti-20S (Enzo Life Sciences; rabbit; 1:2,000; PW9355), anti-Mpk1 (Santa Cruz; mouse; 1:500; sc-374434), anti-p-Mpk1 (Cell Signaling Technology; rabbit; 1:1,000; 9101) and anti-p-Rps6 (Cell Signaling Technology; rabbit; 1:1,000; 2211). Rabbit anti-Adc17 antibody from Bertolotti laboratory was used in all figures except for Figs. 1f and 7f,j and Extended Data Fig. 4b, where sheep anti-Adc17 antibody was used instead owing to stock availability. Anti-mouse IgG, HRP-linked antibody (Cell Signaling Technology; 1:10,000; #7076) and anti-rabbit IgG, HRP-linked antibody (Cell Signaling Technology; 1:10,000; #7074).

SDS-PAGE was employed to resolve the proteins which were transferred to the nitrocellulose membrane using the Tris–Cl (pH 7.5), glycine, methanol and SDS-based transfer buffer. After the transfer, the membrane was blocked with 2.5% bovine serum albumin (BSA) (catalogue no.: MB083; HiMedia). The membrane was incubated in the primary antibody for 90 min and then washed. Next, the membrane was incubated in a secondary antibody for 45 min and washed. Finally, the dried membrane was visualized using LI-COR infrared imaging system. Primary and secondary antibodies used for immunoblotting were as follows: α-TBP1 polyclonal antiserum from rabbit (generated in the laboratory), anti-phospho-p44/42 (Cell Signaling, catalogue no. 4370S), anti-Mpk1 (Santa Cruz Biotechnology, Inc., catalogue no. SC-6803), goat anti-rabbit IgG secondary antibody (catalogue no.: A32734; Invitrogen) and rabbit anti-goat (catalogue no.: A21088; Invitrogen).