Fig 2: NF-κB elements are essential for RelB to enhance Tat recruitment to the LTR. a–d HeLa cells (1 × 107) were transfected with pHIV-1-LTR-luc (6 μg) or pHIV-1-LTR mutNF-κB (6 μg), together with pFlag-Tat (6 μg) or vector plasmids. The relative expression of Tat and RelB in the transfected cells was monitored by western blots (a). Chromatin of the fixed cells was immunoprecipitated with the indicated antibodies. Samples were assessed for enrichment of HIV-1 LTR DNA by PCR (b). Tat (c) and RNA Pol II (d) associated HIV-1 LTR DNA was quantified by real-time PCR. (E to H) Control and RELB−/− HeLa cells (1 × 107) were transfected with pHIV-1-LTR mutNF-κB (6 μg), together with pFlag-Tat (6 μg) or vector plasmids (e). Samples were assessed for enrichment of HIV-1 LTR DNA by PCR. f Tat (g) and RNA Pol II (h) enriched HIV-1 LTR DNA was quantified by real-time PCR

Fig 3: Endogenous RelB facilitates Tat-induced, TAR-independent HIV-1 LTR transactivation. a Control and RELB−/− HeLa cells (0.1 × 106) were transfected by indicated plasmids (pHA-Tat: 100 ng, pHA-RelB: 100 ng) and pHIV-1-LTR-luc luciferase reporter plasmids (100 ng). b Schematic representation of the CycT1-Tat-TAR ternary complex and TAR mutation. c Control and RELB−/− HeLa cells (0.1 × 106) were transfected with the indicated plasmids (pHA-Tat: 100 ng, pHA-RelB: 100 ng) and pHIV-1-LTR-luc luciferase reporter plasmids (100 ng). To normalize transfection efficiency, phRluc-TK (1 ng) was co-transfected. The Rel. Luc. Act in the figure was calculated relative to normalized luciferase activity of pHIV-1-LTR-luc with Tat (with or without RelB) and vectors co-transfected cells. The relative expression of RelB in the transfected cells was monitored by western blotting

Fig 4: The effect of HZ08 on phosphorylation of PI3K, Akt and IKKα. a and b, PC-3 cells were treated with HZ08 alone (a) or treated with HZ08 and IR (b). Cellular extracts from the treated cells were used to quantify the levels of PI3K, p-PI3K, Akt, p-Akt, IKKα, p-IKKα by Western blots. c and d, PC-3 cells were pretreated with LY294002 (25 μM), HZ08 (5 μM) and treated with 6 Gy IR. P-Akt, MnSOD in cellular extracts and RelB in nuclear extracts were quantified by Western blots (c). Cytotoxicity was analyzed using MTT by three independent experiments and plotted as mean ± SD. ** (P < 0.01) shows the significances between two groups as indicated (d)

Fig 5: TP inhibited the BDL-induced ductular reaction by reducing the expression of RelB in cholangiocytes, thereby alleviating liver injury, fibrosis, and inflammation. Created with BioRender.com.