Fig. 6.

RASSF1A regulates proliferation and glycolytic metabolism in IPAH-PASMCs. a Pulmonary vessels from frozen lung sections of a, upper IPAH patients (n = 9), a, lower COPD-PH (n = 8) patients and donors (n = 8–9) were collected via laser-assisted microdissection. RNA was isolated and real time PCRs for indicated genes were performed. b, c Representative paraffin lung tissue sections from donors, IPAH patients, and COPD-PH patients were subjected to immunofluorescence staining of RASSF1, HIF1α (HIF1A), alpha smooth muscle actin (α-Actin), von-willebrand factor (vWF) and collagen 1 (Col1). Nuclei are counterstained with DAPI (blue). Scale bar: 20 μm. d, e Expression of RASSF1A in IPAH- vs donor-PASMCs as analyzed using d real time PCRs and e, upper western blotting, followed by e, lower densitometric quantification of relative RASSF1A expression. f Human PASMCs from IPAH patients were transfected with RASSF1 siRNA (si-RASSF1) and control siRNA (si-Control). 6 h after transfection, cells were placed in medium with growth factors for 48 h. Proliferation was measured by BrdU incorporation assay. g, h Cells were treated with siRNA as above-mentioned and cell lysates were subjected to g real time PCRs and h, left western blotting, followed by h, right densitometric quantification of relative PDK1, LDHA, and HK2 to ACTB expression. *P < 0.05, **P < 0.01, ***P < 0.001 compared to a–e donor or f–h si-Control, unpaired Student’s t-test. n = 3 independent experiments from 3 biological replicates, Data represent mean ± s.e.m.