Fig. 2: HDAC7 is a key deacetylase that regulates PSAT1 acetylation and protein stability.

a–d H1299 cells were transfected with Flag-PSAT1 plasmid alone or co-transfected with HA-HDAC4 (a), HA-HDAC6 (b), HA-HDAC7 (c), or HA-HDAC10 (d). The immunoprecipitations were blotted with anti-HA or anti-Flag antibodies. e, f H1299 cells were transfected with HA-HDAC4 or HA-HDAC7 plasmid alone or co-transfected with Flag-PSAT1. The immunoprecipitations were blotted with anti-HA or anti-Flag antibodies. g In H1299 cells, Flag-PSAT1 and HA-HDAC4 or HA-HDAC7 plasmids were co-transfected. Co-IP was used to detect the ubiquitination level of PSAT1. h, i In H1299 cells, Flag-PSAT1 and HA-HDAC4 or HA-HDAC7 plasmids were co-transfected. The acetylation of PSAT1 was detected. j H1299 cells were transiently transfected with CTL siRNA or HDAC7 selective siRNAs. The acetylation of PSAT1 was detected. k, l In H1299 cells, endogenous interaction between PSAT1 and HDAC7 was tested using anti-Flag and anti-HA antibodies for Co-IP. The immunoprecipitations were blotted with anti-HDAC7 or anti-PSAT1 antibodies. m, n In H1299 cells, HDAC7 was overexpressed or knocked down and to detect PSAT1 protein by Western blot. The relative PSAT1 expression compared with that of Tubulin was quantified. Data represented as the average of three independent experiments (mean ± SD), **P < 0.01, ***P < 0.001. o, p HDAC7 was overexpressed or knocked down in H1299 cells. Co-IP was used to detect the ubiquitination level of PSAT1. q, r H1299 cells were experimented with CHX (100 μg/mL) for various time points and were transfected with HA-HDAC7 or HDAC7 selective siRNA. The PSAT1 protein expressions were detected by Western blot. The relative PSAT1 expression compared with that of Tubulin was quantified. Data represented as the average of three independent experiments (mean ± SD), **P < 0.01, ***P < 0.001.