Fig 1: Inhibition of HIV expression by Nurr1 agonist 6-MP.A, Representative flow cytometry histograms showing the impact of Nurr1 agonist 6-MP on HIV expression in HC69 cells in a dose-dependent manner. B, Western blot detection of Nur77, Nurr1, Nor1, HIV-1 Nef protein, and Nurr1 target gene MMP2 in HC69 cells. Proteins were detected using a rabbit polyclonal anti-Nurr1 antibody (Sata Cruz Biotechnology, Cat# sc-991), a mouse monoclonal anti-Nor1 antibody (Perseus Proteomics, Cat# PP-H7833-00), a rabbit polyclonal anti-Nur77 antibody (Cell Signaling, Cat# 3960S), a mouse monoclonal anti-HIV Nef antibody (Abcam, Cat#ab42355), and a rabbit monoclonal anti-MMP2 antibody (Cell Signaling, Cat#40994). The level of ß-tubulin was used as a loading control. 6-MP at 5 µM strongly induced Nur77 expression, which is consistent with a previous report [123] but did not effect Nurr1 or Nor1 expression. C, Western blot detection of Nef expression from individual clones derived from HIV-infected immortalized human microglial cell (C20) [18] using the pHR’ HIV reporter. The right panel shows the parental mixed population of HIV infected C20 cells. Cells were cultured in the presence of DMSO (placebo) or 6-MP (1 µM) for 48 hr. ß-Tubulin was used as a loading control. D, Western blot measurement of Nef protein from HC69 cells that were untreated or induced with LPS (1 µg/ml), IL-1ß (50 pg/ml), and poly (I:C) (PIC, 500 ng/ml) for 24 hr. The cells were chased in the absence or presence of 1 µM 6-MP for 72 hr after stimulation. ß-Tubulin was used as loading control.