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. 2020 May 13;6(20):eaaz8411. doi: 10.1126/sciadv.aaz8411

Fig. 1. LEDGF/p75 is required for proviral transcription during HIV latency and proviral reactivation.

Fig. 1

(A) RT-qPCR and Western blot analysis of the knockdown efficiency of LEDGF/p75 short hairpin RNAs (shRNAs) in E4 cells 72 hours after lentiviral transduction. Data are represented as means ± SD (n = 3). (B) Flow cytometry analysis of GFP+ cells at different time points post-LEDGF/p75 depletion in E4 cells. A representative flow histogram and quantification of GFP+ cells (means ± SD; n = 3) are shown. FITC, fluorescein isothiocyanate. (C) RT-qPCR analysis of GFP transcript level in E4 cells depleted for LEDGF/p75 for different days. Transcript was normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and shown as means + SD (n = 3). (D) Effect of LEDGF/p75 depletion on HIV proviral reactivation by flow cytometry analysis. A representative flow histogram of HIV proviral reactivation and their quantification (means ± SD; n = 2) are shown. (E) LEDGF/p75 deficiency restrains HIV reentry into latency. Cells were fully activated with TNFα overnight, and the percentage of GFP+ cells was monitored by flow cytometry at indicated time points after TNFα withdrawal. The percentage of GFP+ cells was quantitated and shown as means + SD (n = 2). *P < 0.05, **P < 0.01, and ***P < 0.001.