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KSHV-induced ligand mediated activation of PDGF receptor-alpha drives Kaposi's sarcomagenesis

Fig 4

KSHV vGPCR can activate PDGFRA by upregulation of its ligands PDGFA/B in mECK36 cells.

(A) mRNA levels determined by RT-qPCR of angiogenesis factors, cytokines and REDOX genes in Tetracycline-inducible vGPCR (TET-vGPCR) and control mECK36 stimulated with doxycycline for 24 hours. Data were from three independent experiments carried out in triplicate and are presented as means ± SD. *P < 0.05. (B) and (C) PDGFB-Luc promoter activity in 293T cells co-transfected with empty vector (control), vGPCR, and either a constitutively activated Rac1 (RacQL) construct (B) or a dominant negative Rac1 (RacN17) (C). Data were from three independent experiments carried out in triplicate and are presented as means ± SD. *P < 0.05. (D) Western blot analysis for PDGFB of TET-vGPCR mECK36 cells induced with doxycycline for 24 hrs in the presence of Rac1 inhibitor EHT1864, the NOX inhibitor DPI, or the ROS scavenger NAC. (E) Phosphorylated and total PDGFRA levels of mECK36 cells stimulated with conditioned media from TET-vGPCR mECK36 cells induced with DOX in the presence or the absence of Rac1, NOX and ROS inhibitors. (F) PDGFB mRNA and vGPCR levels of cells in (D) determined by RT-qPCR. Data were from three independent experiments carried out in triplicate and are presented as means ± SD. *P < 0.05.

Fig 4

doi: https://doi.org/10.1371/journal.ppat.1007175.g004