Fig. 4: ACSS2 regulates PPARγ transcriptional activity by SIRT1.

A Representative immunoblot of exogenous IP of HA-tagged PPARγ, HDAC3, Sin3A, NCoR1 and ACSS2 in HEK293T cells (n = 2 biological replicates). B Representative immunoblot of exogenous IP of HA-tagged PPARγ, HDAC3, NCoR1 and ACSS2 following transfection with PPARγ, ACSS2 and SIRT1 or small RNA interference against ACSS2 in HEK293T cells (n = 2 biological replicates). C Representative immunoblot of IP of PPARγ, Sirt1, Sin3A and NCoR1 in the differentiated beige adipocytes from wild type or Asss2^−/− mice (n = 2 biological replicates). D Representative immunoblot of exogenous IP of HA-tagged PPARγ, SIRT1 and ACSS2 in HEK293T cells (n = 2 biological replicates). E Representative immunoblot of exogenous IP of HA-tagged PPARγ, ACSS2 and acetylated proteins in HEK293T cells (n = 3 biological replicates). F Representative immunoblot of adipocyte endogenous PPARγ acetylations in the differentiated beige adipocytes from wild type or Asss2^−/− mice (n = 2 biological replicates). G Representative immunoblot of exogenous IP of ACSS2 and SIRT1 in HEK293T cells (n = 3 biological replicates). H, I Representative immunoblot of exogenous IP of HA-tagged PPARγ, SIRT1, P300 and ACSS2 in HEK293T cells (n = 3 biological replicates). J, K Representative immunoblot of exogenous IP of HA-tagged PPARγ (WT), its deletion mutant or acetylation mutants with ACSS2 in HEK293T cells (n = 2 biological replicates). L, M Representative BiFC images of HEK293T cells transfected with 2 μg plasmid encoding ACSS2 and PPARγ WT, LBD deletion mutant ΔLBD, DBD deletion mutant ΔDBD, K238Q, K238R, K263Q, K263R K154Q, K154R, K155Q or K155R mutants fused to the fluorescent protein fragments indicated in each panel (n = 2 biological replicates). N Representative immunoblot of exogenous IP of P300 and ACSS2 in HEK293T cells transfected with wild type PPARγ and its acetylation mutants (n = 2 biological replicates). O Representative immunoblot of exogenous IP of PPARγ, ACSS2 and PRDM16 in HEK293T cells (n = 2 biological replicates). P Representative immunoblot of exogenous IP of PPARγ, SIRT1 and ubiquitinated protein or (Q) PPARγ, ACSS2 and ubiquitinated protein in HEK293T cells (n = 2 biological replicates). R Representative immunoblot of adipocyte endogenous PPARγ ubiquitinations in the differentiated beige adipocytes from wild type or Asss2^−/− mice (n = 2 biological replicates). S Model of SIRT1 recruitment by ACSS2 to regulate PPARγ transcriptional activity homeostasis. P300-meidated ACSS2 recruitment to acetylated PPARγ can facilitate corepressors HDAC3, Sin3A and NCoR1 disassociation and SIRT1 loading. SIRT1 arrival deacetylated PPARγ, also trans-located ACSS2 from PPARγ to P300 to prevent its over-activation. Besides, SIRT1-mediated PPARγ deacetylation triggers its ubiquitination and degradation. In this scenario, ACSS2 controls PPARγ transcription activity through SIRT1.