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. 2023 Oct 1;37(19-20):865–882. doi: 10.1101/gad.350736.123

Figure 2.

Figure 2.

Characterization of MYC acetylation, stability, and stimulation of cell proliferation in MYC transformed Rat1a fibroblasts. (A) Western blot analysis of total MYC in WCEs of control (E) and the indicated MYC-overexpressing Rat1a cell lines. (Vinc) Vinculin. Ectopic FLAG-MYC and endogenous MYC are indicated. (B) Rat1a-MYC cell lines were treated with cycloheximide (CHX) for the indicated times, and WCEs were analyzed by Western blot. (C) Rat1a-E (control) and RAT1a-MYC/WT cell lines were treated for 2 h with the indicated inhibitors before WCE and IP analyses by Western blot with MYC and MYC AcK site-specific antibodies. (D) Western blot analyses of WCEs and anti-FLAG immunoprecipitated material from the indicated cell lines. (E) C646 inhibition of MYC acetylation analyzed by Western blot, as above. (F) Western blot analyses of the stability of total and site-specific acetylated MYC in Rat1a-MYC/WT cells treated with HDAC inhibitors for 2 h, followed by CHX (in the presence of HDAC inhibitors) for the indicated times. (G) Proliferation of Rat1a-E and Rat1a-MYC (WT or R mutants) cell lines on normal adherent cell culture plates.