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. Author manuscript; available in PMC: 2021 Nov 21.
Published in final edited form as: Sci Immunol. 2021 May 21;6(59):eabe1935. doi: 10.1126/sciimmunol.abe1935

Figure 2. Transcription elongation complex is associated with HDAC 8/9 and is required for MAP2K3 gene expression following HDAC inhibition.

Figure 2.

(A) Proteins identified by mass spectrometry of nHEK extracts enriched by immunoprecipitation with anti-HDAC8 or HDAC9 compared with control IgG. (B) The common enriched nucleosome proteins precipitated by anti-HDAC8 and HDAC9. (C) Transcriptional elongation FACT proteins SSRP1 and SUPT16H that were identified in B were silenced in nHEKs and then HDAC activity chemically inhibited by butyrate treatment (2 mM) for 1 hour. nHEKs were subsequently cultured with or without poly I:C (1 μg/ml) for 4 hours and gene expression measured by qPCR. (N=3; one-way ANOVA) (D) MAP2K3 transcriptional elongation assay. The transcriptional elongation rate of MAP2K3 was measured after SSRP1 or SUPT16H silencing with or without butyrate treatment (2mM) for 1 hour. Data show relative abundance of pulse-labeled RNA for indicated genes at the indicated times after removal of transcriptional inhibitor 6-Dichlorobenzimidazole 1-β-d-ribofuranoside (DRB). (N=3; one-way ANOVA). The results are expressed as the mean ± standard deviation (SD). *, P < 0.05. Data are representative of three independent experiments.