Gesicle Treatment Reduces HIV Proviral Activity
(A) Cell populations treated with LTR gRNA 2 and LTR gRNA 4 were stimulated with pro-inflammatory factors, and cell lysates were taken for the luciferase assay. LTR gRNA 2 and 4 significantly decreased proviral activity after stimulation with TNF-α (50 ng/mL). Additionally, LTR gRNA 4 significantly decreased proviral activity under both basal conditions and LPS (100 ng/mL) stimulation. (C) Lysates were also run on a WES and probed for the HIV protein Nef. (B) Densitometry analysis confirms that HIV viral protein Nef is decreased using LTR gRNAs 2 and 4 after TNF-α treatment. (D) Immunohistochemistry also showed decreased immunoreactivity of Nef in cells treated with LTR gRNA 4. Data are the mean ± SEM of two stable cultures, with three experiments each, analyzed using a two-way ANOVA with post hoc test for each stimulation group (A) or one-way ANOVA with post hoc test (B). *p < 0.05 versus untreated cells: unstimulated; #p < 0.05 versus untreated cells: LPS stimulated; ˆp < 0.05 versus untreated cells: TNF-α stimulated. 2, LTR gRNA 2; 4, LTR gRNA 4; P, protamine; U, untreated.