HIV-1 Vpr involves C/EBP-δ and AP-1 transcription factors in the induction of IL-6 and IL-8 in astrocytes

2015-08-13T03:02:47Z (GMT) by Mohitkumar R. Gangwani Anil Kumar
<p>. SVGA astrocytes were cultured and seeded in 6 well plates. The cells were transfected with siRNA against C/EBP isoforms and AP-1 for 48h. Then the cells were transfected with a plasmid encoding HIV-1 Vpr or were mock-transfected. The cells were harvested at 6h post-transfection followed by the determination of IL-6 and IL-8 mRNA expression levels using real-time RT-PCR. The cell culture supernatants were collected at 48h post-transfection, and protein concentration of secreted IL-6 and IL-8 was determined using BioPlex multi-cytokine assay. <b>(A)</b> Depicts the effect of siRNA against p38β on NF-κB p65 nuclear translocation. <b>(B, D)</b> mRNA expression levels of IL-6 and IL-8 calculated relative to mock-transfected controls after siRNA mediated silencing, respectively. <b>(C, E)</b> Protein concentrations for secreted IL-6 and IL-8 after siRNA mediated silencing, respectively. <b>(F, G)</b> Depicts the effect of siRNAs against p38β and p38δ on the expression levels of C/EBP-δ and p-c-jun following Vpr transfection of astrocytes, respectively. Every bar represents the mean ± SE of thee independent experiments done in triplicates. Statistical analyses were performed using 1-way ANOVA using post-hoc Tukey HSD test, <b>#</b> p < 0.01, <sup><b>a</b></sup> p < 0.05 as compared to Vpr transfected cells; <b>**</b> p < 0.01 and <b>*</b> p < 0.05 compared to mock-transfected controls.</p>