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Fig. 4 | Epigenetics & Chromatin

Fig. 4

From: Interplay between FACT subunit SPT16 and TRIM33 can remodel chromatin at macrophage distal regulatory elements

Fig. 4

Nucleosome dynamics and non-coding RNA production at the − 35 kb regulatory region. a TRIM33, SPT16 and RNA Pol II occupancy along with ChromHMM analysis over the Atp1b3 − 35 kb site. b Nascent bidirectional transcripts at the − 35 kb site in WT and Trim33−/− BMDM, measured after 4sU-tagging and purification of newly transcribed RNA. Data are relative to WT BMDM. Mean ± SEM, n = 3. c Nucleosome positioning at the − 35 kb region in MNase-digested chromatin from WT and Trim33−/− BMDM. Data are relative to the undigested chromatin. Mean ± SEM, n = 3. d ChIP-qPCR of SPT16, PU.1 and TRIM33 occupancy at the − 35 kb site in WT BMDM treated with DMSO or CBL for 4 h. Mean ± SEM, n = 3. e Kinetics of non-coding transcripts at the − 35 kb region in WT and Trim33−/− BMDM after CBL treatment. RT-qPCR was performed using total RNA, and data are presented relative to expression of DMSO-treated WT BMDM. Mean ± SEM, n = 3. f SPT16 ChIP-qPCR (left) and nucleosome positioning (right) at the − 35 kb region in Trim33−/− IM (Trim33−/−) and in Trim33−/− IM that contained a lentivirus expressing TRIM33 (Trim33−/− + WT TRIM33). Mean ± SEM, n = 3. g Non-coding transcripts at the − 35 kb region (left), Atp1b3 mRNA (middle) and protein expression (right) in Trim33−/− and Trim33−/− + WT TRIM33 IM. Data are presented relative to expression of Trim33−/− IM. Mean ± SEM, n = 4. h Model for FACT/TRIM33-mediated repression at the Atp1b3 locus

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