Fig. 5: Tip60 acetylates eEF1A1 in a Stat3-dependent manner.

a, e eEF1A1 immunoprecipitation (IP) in lysates of SCs induced to de-differentiate for 1 day and treated with TH1864 (Tip60 inhibitor, Tip60i) or vehicle, and Tip60, Stat3 (a) or Ac-eEF1A (e) Western blot and quantification (normalized to eEF1A1) of Tip60 co-immunoprecipitated with eEF1A1 (a) or of Ac-eEF1A1 (e) in Tip60 inhibitor- compared to vehicle-treated SCs. b IP Stat3 or Flag (Ctrl) in lysates of SCs induced to de-differentiate for 1 day, Tip60 Western blot and Stat3 inputs. c Tip60, GAPDH (cytoplasmic marker) and Lamin A/C (nuclear marker) Western blots in cytoplasmic and nuclear fractions of SCs induced to de-differentiate for 1 day. Dashed lines: samples run on the same gel, but not on consecutive lanes. d Tip60 and GAPDH Western blots in lysates of crushed (Cr) and contralateral (Co) mouse sciatic nerves at 1-3-5-12dpl, and Tip60 quantification normalized to GAPDH in Cr compared to Co. f Sox10 and GAPDH Western blots in lysates of SCs induced to de-differentiate and treated with Tip60i or vehicle for 3 days and Sox10 quantification normalized to GAPDH. g Stat3 and Tip60 Western blots in SCs transduced with Stat3-specific shRNA (Stat3 sh) or non-targeting control shRNA (Ctrl sh) lentiviruses and quantification of sumoylated (higher molecular weight) and non-sumoylated (lower molecular weight) Tip60 isoforms normalized to GAPDH. h eEF1A1 IP and Tip60 Western blot in SCs transduced with Stat3 sh or Ctrl sh lentiviruses. i Stat3 and Ac-eEF1A co-immunofluorescence and DAPI labeling in primary SCs transduced with Stat3 sh or Ctrl sh lentiviruses and induced to de-differentiate for 2 days. Scale bar: 20 µm. Data presented as mean values ± SEM. Unpaired (f) or paired (a, d, e, g) one-tailed (gray asterisk) or two-tailed (black asterisks) Student’s t-tests. P values = 0.029197 (a), 0.139771 (d, 1dpl), 0.047935 (d, 3dpl), 0.030298 (d, 5dpl), 0.061845 (d, 12dpl), 0.001796 (e), 0.033551 (f), 0.047979 (g, sumoylated), 0.300207 (g, non-sumoylated). N = 3 independent experiments or 3 animals per time-point. For each panel, representative images of 3 independent experiments are shown. Source data are provided as a Source Data file.