Kaposi’s Sarcoma-Associated Herpesvirus Subversion of the Anti-Inflammatory Response in Human Skin Cells Reveals Correlates of Latency and Disease Pathogenesis
Figure 7
A link between KSHV latency and the MC1-R signaling axis in skin-derived cell lines. (a) Western blot analysis of phosphorylated NF-κB p65, MC1-R, and TRP-1 in total cell lysates extracted from MeWo (a) or Mel1700 (b) cells either uninfected (control) or acutely infected with KSHV for 0.3 h, 1 h, 3 h, or 6 h. GAPDH was used as loading control. (c) ImageJ quantitation of the p65 band intensities in (a) and (b) relative to GAPDH controls. (d) Mel1700 cells were infected in 6-well plates with increasing volumes (mL/well) of concentrated supernatant containing infectious KSHV, and total RNA from infected cells was subjected to RT-PCR using primer sets for host anti-inflammatory MC1R, POMC, and SLC7A11. (e) Equal aliquots from the same RNA used in (d) were subjected to RT-PCR analysis for select viral latency and cell growth control genes (i.e., GPCR, LANA, and v-FLIP). (f) Western blot analysis of the melanoma cell marker, Melan A, and anti-inflammatory genes MC1-R, TRP1, and SLC7A11 in total cell lysates of uninfected (−) or chronically infected (+) long-term cultures of MeWo-KSHV and Mel1700-KSHV cells. GAPDH was used as an internal control for both the RT-PCR (e) and western blot assays.
