Pfi 1

Apabetalone (RVX-208; BET inhibitor, S7295), ARV-825 (BRD4 specific inhibitor, S8297), AZD-5153 6-hydroxy-2-naphthoic acid (BET/BRD4 inhibitor, S8344), CPI-203 (BET inhibitor, S7304), Molibresib (I-BET-762; BET inhibitor, S7189), (+)-JQ1 (BET inhibitor, S7110), INCB054329 (BET inhibitor, S8753), MS436 (BET inhibitor, S7305), Birabresib (OTX015; BET inhibitor, S7360), PLX51107 (a new BET inhibitor, S8739), and PFI-1 (PF-6405761; BRD2/BRD4 inhibitor, S1216) were purchased from Selleck.cn, and ZL0580 (a BRD4-specific inhibitor) was prepared as previously described (20 (link), 45 (link)). The structures and functions of these inhibitors are shown in Fig. 3A, Text S1 in the supplemental material (see also Selleck.cn [https://www.selleck.cn]), and Table 1 (20 (link), 41 (link), 46 (link)– (link)55 (link)).

J-Lat C11 cells^{24 (link),26 (link)} (established in our lab) and A10.6 cells^{29 (link),54 (link)} (obtained from NIH AIDS Reagent Program) harboring latent, transcriptionally competent HIV-1 provirus that encodes GFP as an indicator of viral activation were cultured in RPMI1640 medium (Gorning) supplemented with 10% fetal bovine serum (FBS) (Gibco), 100 U/mL penicillin and 100 μg/ml streptomycin (Gibco) in a 37 °C incubator containing 5% CO₂. For in vitro reactivation experiments, C11 cells or A10.6 cells were stimulated with RVX-208 (Selleckchem), PFI-1 (Selleckchem), JQ1 (Selleckchem), SAHA (Sigma-Aldrich), prostratin (Sigma-Aldrich) or TNF-α (Sigma-Aldrich) respectively and subjected to determine GFP expression using flow cytometry (BD Calibur).

MCF7 cancer cells were seeded at 5 × 10⁵ in a six-well plate overnight. The next day, cells were treated with 200 nM to1 μM JQ1 dissolved in DMSO, 500 μM PFI-1 (Selleckchem) dissolved in DMSO or 100 μM I-BET151 (Selleckchem) dissolved in EtOH. The following day, MCF7 cells were washed and the effect on CD47 expression was analysed by qPCR over a period of 1 week for JQ1 and 2 days for PFI-1 and I-BET151. Jurkat cancer cells were treated with JQ1 following the same procedure performed for MCF7 cells. Jurkat cells were washed the next day after JQ1 treatment and effect on CD47 expression analysed by qPCR over a period of 48 h. For this experiment, we used DMSO as a vehicle.