Fig 2: Recruitment of STAT5 and CBP/P300 to the LTR by benzodiazepines in TZMBL Cells. TZMBL cells were treated with Benzodiazepines: RO5-3335, Alprazolam and Clonazepam with or without HDACi SAHA for 48 hrs at the indicated concentrations. (A) Cells were lysed and LTR driven luciferase was determined. (B) ChIP analysis was performed to determine RUNX1 binding to the HIV-1 LTR. (C) ChIP analysis was performed to determine STAT5 and CBP/P300 binding to the HIV-1 LTR. **P = 0.01, ***P = 0.001, ****P = 0.0001, ns = not significant.

Fig 3: A proposed model of regulation of the IFN-JAK/STAT1 signaling by USP12.USP12 translocates from cytoplasm to nucleus in IFN signaling. The decrease in cytoplasmic USP12 levels facilitates CBP-mediated acetylation and activation of IFN signaling. USP12 accumulation in the nucleus maintains activated p-STAT1 levels by blocking CBP-mediated acetylation of p-STAT1 and subsequent TCPTP-mediated dephosphorylation of p-STAT1, which enables efficient IFN antiviral response.

Fig 4: Recruitment of STAT5 and CBP/P300 to the LTR in Primary Cells with RUNX1 binding inhibition. PBMC blasts from healthy donors were infected with ?Nef or ?RUNX. (A) Viral replication was followed over time by p24 ELISA. After 48 h infection, PBMC cells were fixed and prepped for CHIP analysis with IP targeting (B) RUNX1 (C) STAT5 and (D) CBP/P300. **P = 0.01, ***P = 0.001.

Fig 5: USP12 translocates from cytoplasm to nucleus in IFN-I signaling.(A) Western blot analysis of USP12 protein levels in the cytoplasm and nucleus of RAW264.7 cells treated with mouse mIFNß (500 IU/ml) for 0, 1 and 3 hrs. (B and C) Western blot analysis of USP12 protein levels in the cytoplasm and nucleus of HEK293T (B) or HCT116 (C) cells treated with IFNa (1,000 IU/ml) for 0, 1.5, 3 and 4.5 hrs. (D) Western blot analysis of HA-USP12 levels in the nucleus of HEK293T cells transfected with HA-USP12 and then treated with IFNa (1,000 IU/ml) as indicated. (E) HeLa cells were treated with IFNa (3,000 IU/ml) for 6 hrs. Cellular CBP and USP12 proteins were stained by specific antibodies, and cell nuclei were stained by DAPI. The fluorescent images were captured with the Nikon A1 confocal microscope.