Skip to main content
. 2016 Jun;6(2):202–210. doi: 10.1086/686012

Figure 3.

Figure 3

Hypoxia-induced proline-rich tyrosine kinase 2 (Pyk2) activation mediates nuclear factor-kappa B (NF-κB) transcriptional activity and NADPH oxidase 4 (Nox4) protein expression in human pulmonary artery smooth muscle cells (HPASMCs). A, HPASMCs were transfected with firefly luciferase vector containing 5 consensus NF-κB binding sites, along with a renilla luciferase vector to normalize for transfection efficiency. Cells were exposed to hypoxia or normoxia for 72 hours and treated with tyrphostin A9 (TA9) or vehicle during the final 24 hours of exposure. Cell lysates were prepared and analyzed for firefly luciferase activity. Each bar represents mean ± SEM firefly luciferase activity/renilla luciferase activity, expressed as fold change versus control; n = 4. *P < 0.05 versus normoxia + TA9. #P < 0.05 versus hypoxia + TA9. B, HPASMCs were transfected with 50 nM control siRNA (Con) or Pyk2 siRNA (Pyk2) and exposed to normoxia or hypoxia for 72 hours. Cell lysates were immunoblotted with an anti-Nox4 antibody. GAPDH (glyceraldehyde 3-phosphate dehydrogenase) levels were used for normalization. Each bar represents mean ± SEM Nox4/GAPDH levels, expressed as fold change versus control; n = 3. **P < 0.01 versus normoxia + Con. ##P < 0.01 versus hypoxia + Con. siRNA: small interfering RNA.