Journal: Respiratory Research
Article Title: TRB3 mediates vascular remodeling by activating the MAPK signaling pathway in hypoxic pulmonary hypertension
Figure Lengend Snippet: Inhibitors of ERK, JNK, and p38 MAPK reversed the effect of TRB3 overexpression on PASMC biological behavior. A The CCK-8 assay was used to evaluated the proliferation of TRB3-overexpressing cells after treating with U0126, SP600125, and SB203580. B – D Western blot analysis of PCNA expression in TRB3 overexpressing cells incubated with U0126, SP600125, or SB203580 for 12 h. E Cell apoptosis induced by U0126, SP600125 and SB203580 in TRB3-overexpressing cells was evaluated using flow cytometry, and the percentage of early apoptotic (Annexin V+/PI−) and late apoptotic (Annexin V+ /PI+) cells was analyzed. F – H Western blot analysis of the protein expression of PARP, BAX, and Bcl2 in TRB3-overexpressing cells incubated with U0126, SP600125, and SB203580 for 12 h. I Crystal violet staining is presented at × 200 magnification for the Transwell assay and the migrated cells were counted and analyzed. J – L Western blot analysis of MMP9 expression in TRB3-overexpressing cells incubated with U0126, SP600125, and SB203580 for 12 h. Data represent the mean ± SEM (n = 4). *P
Article Snippet: Polyclonal antibodies against PCNA, BAX, caspase 3, PARP, HRP-conjugated β-actin, and Survivin were purchased from Proteintech (USA); anti-Bcl2 antibody was obtained from Boster (China); anti-MMP9 antibody was obtained from Affinity (USA); anti-p38/p-p38 MAPK, anti-JNK/p-JNK, anti-ERK/p-ERK, and normal rabbit IgG antibodies were acquired from Cell Signaling Technology (USA); normal mouse IgG was obtained from Santa Cruz Biotechnology (USA).
Techniques: Over Expression, CCK-8 Assay, Western Blot, Expressing, Incubation, Flow Cytometry, Staining, Transwell Assay