Fig 1: Schematic illustration indicating FNDC5-deficient mice experienced bone loss by inhibiting osteogenic differentiation and promoting osteoclast activation. (A) Schematic diagram of the mechanism of irisin regulating bone formation. Irisin combined with its receptor integrin αV on the surface of BMSCs, then in turn activated the phosphorylation of ERK and STAT3, promotes an increase of BMP2. It banded to the receptor BMPR2 on the membrane surface to activate the phosphorylation of Smad1/5/9 and promote osteogenic differentiation.
Fig 2: The lncRNA MEG3 promotes trophoblast cell metastasis and invasiveness through its regulation of the miR-21/BMPR2 signaling pathway. A,B) RT-qPCR analysis of the transcription levels of miR-21-5p and MEG3 in the placenta for the normal pregnancy group and PE patients. C) Correlation analysis between the transcription levels of miR-21-5p and MEG3 in the placental tissues of PE patients; *p<0.05 relative to the normal pregnancy group. Data are presented as the means ±SEM, n = 3.
Fig 3: The effect of OIP5 knockdown on downstream genes. (A) Analysis of downstream genes of OIP5 using IPA analysis. (B) RT-qPCR was used to determine the changes in the expression of OIP5 downstream genes in Huh7 cells at 72 h following pGCSIL-shOIP5 transfection. Samples were normalized to GAPDH mRNA expression. RT-qPCR results are indicated as mean ± standard deviation of three independent experiments performed in triplicate. GAPDH protein was used as the internal control. *P<0.05 and **P<0.01 vs. pGCSIL-shCtrl-transfected cells. (C) The expression of OIP5 downstream proteins were analyzed by western blot analysis in Huh7 cells transfected with pGCSIL-shOIP5 lentivirus. #, target genes with a Z-score >3.0; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; Ctrl, control; sh, short hairpin; OIP5, Opa interacting protein 5; mir-1, microRNA1; CDKN2A, cyclin dependent kinase inhibitor 2A; TWF1, twinfilin actin binding protein 1; H3F3A, H3 histone family member 3A; HNRNPU, heterogeneous nuclear ribonucleoprotein U; EEF1A1, eukaryotic translation elongation factor 1a1; RB1, retinoblastoma 1; MAD2L1, mitotic arrest deficient 2 like 1; BTG2, B-cell translation gene 2; BMPR2, bone morphogenetic protein receptor type 2; CHEK1, checkpoint kinase 1; RAC1, Rac family small GTPase 1; PDIA3, protein disulfide isomerase family A member 3; RPS6KB1, ribosomal protein S6 kinase B1; CUL4B, cullin 4B.
Fig 4: Irisin promoted the phosphorylation of ERK/STAT through binding to integrin receptor aV and up-regulates the expression of BMP2 to enhance osteogenic differentiation. (A) Cell lysates were subjected to immunoblotting analysis. Protein expression levels of integrin aV, p-Erk1/2, p-STAT3, BMPR2, p-Smad1/5/9, and Smad4 of BMSCs induced into osteoblasts for 3 days in the presence of r-irisin and SB273005. (B) ALP and ARS staining. (C) Protein expression levels of BMP2, Smad1/5/9, p-Smad1/5/9 and Smad4 after 3 days of osteogenic induction of BMSCs treated with r-irisin, and the ratio of p-Smad1/5/9 to Smad1/5/9 of the protein grayscale values. (D) Immunohistochemical staining of p-Smad1/5/9 in the femur of WT and FNDC5-/- mice (n=6, *P <0.05, ***P <0.001, ****P <0.0001).
Fig 5: Expression assay results of c.419-43delT carriers and wild-type patients. (A) Dot plot representing the fold change between carriers and wild-type patients without taking into consideration cell cycle stage. The results showed a slight increase in the variant carriers’ BMPR2 expression, and these data were not statistically significant. (B) Dot plot representing the fold change between variant carriers and wild-type patients taking into account cell cycle stage. Data showed a rise of gene expression in the G0/G1 and a slight decrease for S phase for the carriers, while in the G2/M, it showed almost identical expression between both groups. There were no statistically significant differences between stages.
Supplier Page from Abcam for Anti-BMPR2 antibody [1F12]