Fig 2: MORC2 activates the PI3K/AKT signaling via binding to NDRG1 promoter in glioma cells. The expression of PTEN, p-PI3K, p-AKT, PI3K and AKT was examined by western blotting. ***P<0.001 vs. pc-NC; ###P<0.001 vs. pc-NDRG1. MORC2, Microrchidia family CW-type zinc finger 2; pc-NC, empty vector; pc-MORC2, overexpression plasmid of MORC2; pc-NDRG1, overexpression plasmid of NRDG1; p, phosphorylated.

Fig 3: PGRMC1 overexpression alters the phosphoproteome and promotes cell survival of normal breast cells.a Time-dependent cell proliferation by MTS assay of PGRMC1-overexpressing MCF10A cells compared with cells transfected with a pCMV empty vector. b Protein levels of normal MCF10A breast epithelial cells transfected with pCMV or PGRMC1-overexpressing plasmid by western blot analysis. c Volcano plots generated from phosphoproteome analysis representing both total and phosphorylated protein forms of MCF10A-overexpressing cells compared with pCMV-transfected MCF10A cells. d, e GO term-enriched pathway analysis by REACTOME and heatmap clusters of differentially expressed genes identified by phosphoproteome analysis following PGRMC1 overexpression in MFC10A cells. f Cluster network analysis of the top 20 differentially expressed genes. g Heat maps depicting increased or decreased phosphorylation at multiple sites of PTEN, AKT, mTOR, P70S6K and EGFR of MCF10A PGRMC1-overexpressing cells. h Protein levels of proliferative markers PTEN, AKT, mTOR, EGFR and their phosphorylation sites in MCF10A pCMV and MCF10A PGRMC1-overexpressing cells by western blot analysis. i Schematic of downstream phosphorylation activity following AG-205 treatment or silencing of PGRMC1 of breast cancer cells. Analysis of a is the mean ± SD of three independent experimental replicates (n = 3). *P < 0.05 calculated using t test comparing pCMV with PGRMC1 OE. Volcano plot: significant down- and upregulated proteins are represented, P < 0.05.

Fig 4: Astrogliosis in the CA3 area of the hippocampus following kainic acid injection.Glial fibrillary acidic protein (GFAP)-positive profiles in the hippocampi of control (A), kainic acid (KA)-treated, 1 day survival mice (B) and KA/bpv(pic)-treated, 1 day survival mice (C) (scale bar: 50 µm). D. Histogram showing the % of GFAP staining in the hippocampal CA3 area one day after kainic acid injection and PTEN inhibition by bpv(pic). The significant increase in GFAP immunoreactivity after kainate administration, expressed as a percentage of the whole area occupied by positive profiles, is significantly prevented by bpv(pic) treatment. *P<0.05, §<0.05.

Fig 5: Pice blocks the PTEN/AKT signaling pathway via inhibiting miR-21 expression in osteosarcoma cells. (A) Saos-2 cells were transfected with miR-21 mimics/mimics NC after treatment with 100 µM of Pice for 24 h, and the expression levels of PTEN, p-AKT and AKT were measured by western blot analysis. ß-actin was used as an internal control. (B) The histogram indicated the relative protein expression obtained for each group. *P<0.05, **P<0.01 vs. control; ##P<0.01. Data were presented as the mean ± standard deviation of three individual experiments. AKT, protein kinase B; miR, microRNA; NC, negative control; p, phosphorylated; PTEN, phosphatase and tensin homolog.