Fig 6.

The NF-κB pathway mediates dexamethasone suppression of IL-1α induced by KSHV miRNAs and vFLIP. (A–C) IL-1α mRNA expression level measured by RT-qPCR in cells harboring wild-type KSHV (KMM) or KSHV with a deletion of the miRNA cluster (ΔmiRNA), vFLIP (ΔvFLIP), or vCyclin (ΔvCyclin) (A), MM cells expressing the miRNA cluster or vector control (B), or vFLIP, vCyclin or vector control (C). (D–F) IL-1Ra mRNA expression level measured by RT-qPCR in KMM cells or cell harboring ΔmiRNA, ΔvFLIP, or ΔvCyclin mutant (D), MM cells expressing the miRNA cluster or vector control (E), or vFLIP, vCyclin or vector control (F). (G) IκBα mRNA expression levels in MM and KMM cells stably expressing IκBαM measured by RT-qPCR. (H) Immunoblots of IκBα, pRelA-S536, and RelA in MM and KMM cells stably expressing IκBαM. (I and J) IL-1α mRNA (I) and protein (J) levels in MM and KMM cells stably expressing IκBαM measured by RT-qPCR and ELISA, respectively. (K) IL-1Ra mRNA expression levels in MM and KMM cells stably expressing IκBαM measured by RT-qPCR. (L) Enrichment plots of TNFA signaling via NF-κB (GSEA hallmark) of MM and KMM cells treated with dexamethasone. (M) IκBα mRNA expression levels in MM and KMM treated with 0 or 0.1 µM dexamethasone for 2 days measured by RT-qPCR. (N) Immunoblot of IκBα of MM and KMM cells treated with 0 or 0.1 µM dexamethasone for 2 days. (O) Immunoblots of pRelA-S536, RelA, p105/p50, and p100/p52 in MM and KMM cells treated with 0 or 0.1 µM dexamethasone for 2 days. Student’s t test was used for the analysis, and results from three independent repeats are presented as mean ± SD (A–G, I, K, and M).