Fig 2: Ctfg suppressed the proliferation and migration in H/R PC12 cells(A) The mRNA expression of Ctgf in H/R PC12 cells transfected with miR-375 mimic and pcDNA3.0-Ctgf or pcDNA3.0-vector was determined by qRT-PCR (B) The effect of miR-375 mimic and pcDNA3.0-Ctgf co-transfection on H/R p12 cells proliferation was detected by EdU assay, with pcDNA3.0-vector as control. (C) The effect of miR-375 mimic and pcDNA3.0-Ctgf co-transfection on H/R PC12 cells’ migration was detected by transwell assay. (D) The effect of miR-375 mimic and pcDNA3.0-Ctgf co-transfection on protein expression of Ctgf, p21, PI3K, and p-AKT in H/R PC12 cells were determined by Western blotting. The experiments were performed in triplicate and each value represents mean ± S.D. *P<0.05 compared with H/R + mimic + pcDNA3.0 vector. **P<0.01 compared with H/R + mimic + pcDNA3.0 vector.

Fig 3: Effects of YAP on cell biological behavior of neuroblastoma cells. (A) Transwell array tested the invasive ability of SK-N-BE (2) and SK-N-AS between siYAP and siNC, and the number of cells crossing the bottom of the chamber were analyzed by GraphPad Prism 5. Scale bar: 100 µm p = 0.0079, respectively. (B) The cell migration ability was estimated using the wound-healing assay after YAP downregulation. The images were captured at the time shown on the graph after wounding (magnification: 50×; scale bar: 250 µm) p = 0.0022, respectively. (C) WB tested the expression of YAP, E-cad, and MMP9 after downregulation of YAP. (D) WB tested the expression of YAP, CTGF, cyclin D1, FGF1, and Prp19 after downregulation of YAP. *p < 0.05 and **p < 0.01.

Fig 4: Norcantharidin (NCTD) sensitizes cells to cisplatin (DDP)-induced tumor growth inhibition. (A) RT-PCR and western blot analysis revealed that the mRNA and protein expression levels of YAP and its targets genes, including CYR61 and CTGF were much higher in the A549/DDP and H1299/DDP cells than in the parent A549 and H1299 cells. (B and C) CCK-8 and MTT assay indicated that a higher, but not lower concentration of NCTD suppressed A549/DDP and H1299/DDP cell proliferation. However, the higher concentration of DDP still did not suppress A549/DDP cell proliferation. (D) Cellular growth and proliferation assay showing that a low dose of NCTD sensitized the A549/DDP and H1299/DDP cells to the anti-proliferative effects of low doses of DDP. Results are presented as the means ± SD, and the error bars represent the SD of 3 independent experiments. *P<0.05; **P<0.01; ***P<0.001 vs. control group. NS, not significant.

Fig 5: Identification of vascular smooth muscle cell clusters under disturbed blood flow.A t-distributed stochastic neighbor embedding (t-SNE) representation of aligned gene expression data in each individual cell extracted from the left carotid artery of wild-type mice 7 days after partial carotid ligation (PCL) or without PCL (control). B t-SNE plot of three VSMCs clusters from the PCL and control. C The heatmap showing differences of biological processes among three distinct VSMCs by GSVA. D Branched pseudotime trajectory, each cell being colored by its pseudotime value (left) and its seurat clusters (right). E The heatmap showing variable genes along the pseudotime trajectory. The X-axis represents cells ordered by pseudotime (from left to right) and different colors correspond to the scaled expression of each gene in each cell (from Monocle). F Representative immunofluorescence microscopy images for TSP-1 and CTGF in the carotid artery from wild-type mice with partial carotid ligation (PCL) or control. bar = 50 µm.