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IRF1 promotes osteogenic differentiation of mBMSCs may through activating PI3K/AKT signaling pathway. a. Venn map revealed the gene expression pattern in OM_IRF1 and OM group. b. Volcano plot displayed the upregulated and downregulated genes in OM_IRF1 group in comparison to the OM group. c. The highest enriched biologic functions were displayed here. d. The highest enriched signaling pathways were displayed here. e.-h. Relative expression of key genes in PI3K/AKT signaling pathway, including Pik3r1 (e), Pik3r2 (f), <t>Akt1</t> (g) and Akt2 (h) measured through RT-PCR (n = 9). i. Protein levels of p-PI3K, PI3K, p-AKT and AKT measured through western blotting analysis (n = 3). j, k. The proportion of p-PI3K (j) and p-AKT (k) were calculated. l. ALP staining and ALP activity quantification of the Irf1 -overexpression mBMSCs and the Irf1 -overexpression mBMSCs with PI3K inhibitor ( LY249002 ) on day 7 (n = 9). m. ARS staining and quantification of the Irf1 -overexpression mBMSCs and the Irf1 -overexpression mBMSCs with PI3K inhibitor ( LY249002 ) on day 14 (n = 9). n. Expression of Alp and Runx2 after 7 days of osteogenic induction by qRT-PCR (n = 9). o. Expression of Spp1 and Bglap after 14 days of osteogenic induction measured by qRT-PCR (n = 9). OM: mBMSCs expressing vector after 7 days of osteogenesis induction; OM_IRF1: mBMSCs over-expressing Irf1 after 7 days of osteogenesis induction. Data are shown as the mean ± SD. *indicates a significant difference from the control group, * P < 0.05, ** P < 0.01, *** P < 0.001.
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IRF1 promotes osteogenic differentiation of mBMSCs may through activating PI3K/AKT signaling pathway. a. Venn map revealed the gene expression pattern in OM_IRF1 and OM group. b. Volcano plot displayed the upregulated and downregulated genes in OM_IRF1 group in comparison to the OM group. c. The highest enriched biologic functions were displayed here. d. The highest enriched signaling pathways were displayed here. e.-h. Relative expression of key genes in PI3K/AKT signaling pathway, including Pik3r1 (e), Pik3r2 (f), <t>Akt1</t> (g) and Akt2 (h) measured through RT-PCR (n = 9). i. Protein levels of p-PI3K, PI3K, p-AKT and AKT measured through western blotting analysis (n = 3). j, k. The proportion of p-PI3K (j) and p-AKT (k) were calculated. l. ALP staining and ALP activity quantification of the Irf1 -overexpression mBMSCs and the Irf1 -overexpression mBMSCs with PI3K inhibitor ( LY249002 ) on day 7 (n = 9). m. ARS staining and quantification of the Irf1 -overexpression mBMSCs and the Irf1 -overexpression mBMSCs with PI3K inhibitor ( LY249002 ) on day 14 (n = 9). n. Expression of Alp and Runx2 after 7 days of osteogenic induction by qRT-PCR (n = 9). o. Expression of Spp1 and Bglap after 14 days of osteogenic induction measured by qRT-PCR (n = 9). OM: mBMSCs expressing vector after 7 days of osteogenesis induction; OM_IRF1: mBMSCs over-expressing Irf1 after 7 days of osteogenesis induction. Data are shown as the mean ± SD. *indicates a significant difference from the control group, * P < 0.05, ** P < 0.01, *** P < 0.001.
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IRF1 promotes osteogenic differentiation of mBMSCs may through activating PI3K/AKT signaling pathway. a. Venn map revealed the gene expression pattern in OM_IRF1 and OM group. b. Volcano plot displayed the upregulated and downregulated genes in OM_IRF1 group in comparison to the OM group. c. The highest enriched biologic functions were displayed here. d. The highest enriched signaling pathways were displayed here. e.-h. Relative expression of key genes in PI3K/AKT signaling pathway, including Pik3r1 (e), Pik3r2 (f), <t>Akt1</t> (g) and Akt2 (h) measured through RT-PCR (n = 9). i. Protein levels of p-PI3K, PI3K, p-AKT and AKT measured through western blotting analysis (n = 3). j, k. The proportion of p-PI3K (j) and p-AKT (k) were calculated. l. ALP staining and ALP activity quantification of the Irf1 -overexpression mBMSCs and the Irf1 -overexpression mBMSCs with PI3K inhibitor ( LY249002 ) on day 7 (n = 9). m. ARS staining and quantification of the Irf1 -overexpression mBMSCs and the Irf1 -overexpression mBMSCs with PI3K inhibitor ( LY249002 ) on day 14 (n = 9). n. Expression of Alp and Runx2 after 7 days of osteogenic induction by qRT-PCR (n = 9). o. Expression of Spp1 and Bglap after 14 days of osteogenic induction measured by qRT-PCR (n = 9). OM: mBMSCs expressing vector after 7 days of osteogenesis induction; OM_IRF1: mBMSCs over-expressing Irf1 after 7 days of osteogenesis induction. Data are shown as the mean ± SD. *indicates a significant difference from the control group, * P < 0.05, ** P < 0.01, *** P < 0.001.
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IRF1 promotes osteogenic differentiation of mBMSCs may through activating PI3K/AKT signaling pathway. a. Venn map revealed the gene expression pattern in OM_IRF1 and OM group. b. Volcano plot displayed the upregulated and downregulated genes in OM_IRF1 group in comparison to the OM group. c. The highest enriched biologic functions were displayed here. d. The highest enriched signaling pathways were displayed here. e.-h. Relative expression of key genes in PI3K/AKT signaling pathway, including Pik3r1 (e), Pik3r2 (f), <t>Akt1</t> (g) and Akt2 (h) measured through RT-PCR (n = 9). i. Protein levels of p-PI3K, PI3K, p-AKT and AKT measured through western blotting analysis (n = 3). j, k. The proportion of p-PI3K (j) and p-AKT (k) were calculated. l. ALP staining and ALP activity quantification of the Irf1 -overexpression mBMSCs and the Irf1 -overexpression mBMSCs with PI3K inhibitor ( LY249002 ) on day 7 (n = 9). m. ARS staining and quantification of the Irf1 -overexpression mBMSCs and the Irf1 -overexpression mBMSCs with PI3K inhibitor ( LY249002 ) on day 14 (n = 9). n. Expression of Alp and Runx2 after 7 days of osteogenic induction by qRT-PCR (n = 9). o. Expression of Spp1 and Bglap after 14 days of osteogenic induction measured by qRT-PCR (n = 9). OM: mBMSCs expressing vector after 7 days of osteogenesis induction; OM_IRF1: mBMSCs over-expressing Irf1 after 7 days of osteogenesis induction. Data are shown as the mean ± SD. *indicates a significant difference from the control group, * P < 0.05, ** P < 0.01, *** P < 0.001.
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IRF1 promotes osteogenic differentiation of mBMSCs may through activating PI3K/AKT signaling pathway. a. Venn map revealed the gene expression pattern in OM_IRF1 and OM group. b. Volcano plot displayed the upregulated and downregulated genes in OM_IRF1 group in comparison to the OM group. c. The highest enriched biologic functions were displayed here. d. The highest enriched signaling pathways were displayed here. e.-h. Relative expression of key genes in PI3K/AKT signaling pathway, including Pik3r1 (e), Pik3r2 (f), Akt1 (g) and Akt2 (h) measured through RT-PCR (n = 9). i. Protein levels of p-PI3K, PI3K, p-AKT and AKT measured through western blotting analysis (n = 3). j, k. The proportion of p-PI3K (j) and p-AKT (k) were calculated. l. ALP staining and ALP activity quantification of the Irf1 -overexpression mBMSCs and the Irf1 -overexpression mBMSCs with PI3K inhibitor ( LY249002 ) on day 7 (n = 9). m. ARS staining and quantification of the Irf1 -overexpression mBMSCs and the Irf1 -overexpression mBMSCs with PI3K inhibitor ( LY249002 ) on day 14 (n = 9). n. Expression of Alp and Runx2 after 7 days of osteogenic induction by qRT-PCR (n = 9). o. Expression of Spp1 and Bglap after 14 days of osteogenic induction measured by qRT-PCR (n = 9). OM: mBMSCs expressing vector after 7 days of osteogenesis induction; OM_IRF1: mBMSCs over-expressing Irf1 after 7 days of osteogenesis induction. Data are shown as the mean ± SD. *indicates a significant difference from the control group, * P < 0.05, ** P < 0.01, *** P < 0.001.

Journal: Journal of Advanced Research

Article Title: IRF1 regulates apoptosis and osteogenic differentiation of bone marrow mesenchymal stem cells and ameliorates osteoporosis by activating the PI3K/AKT signaling pathway

doi: 10.1016/j.jare.2025.11.037

Figure Lengend Snippet: IRF1 promotes osteogenic differentiation of mBMSCs may through activating PI3K/AKT signaling pathway. a. Venn map revealed the gene expression pattern in OM_IRF1 and OM group. b. Volcano plot displayed the upregulated and downregulated genes in OM_IRF1 group in comparison to the OM group. c. The highest enriched biologic functions were displayed here. d. The highest enriched signaling pathways were displayed here. e.-h. Relative expression of key genes in PI3K/AKT signaling pathway, including Pik3r1 (e), Pik3r2 (f), Akt1 (g) and Akt2 (h) measured through RT-PCR (n = 9). i. Protein levels of p-PI3K, PI3K, p-AKT and AKT measured through western blotting analysis (n = 3). j, k. The proportion of p-PI3K (j) and p-AKT (k) were calculated. l. ALP staining and ALP activity quantification of the Irf1 -overexpression mBMSCs and the Irf1 -overexpression mBMSCs with PI3K inhibitor ( LY249002 ) on day 7 (n = 9). m. ARS staining and quantification of the Irf1 -overexpression mBMSCs and the Irf1 -overexpression mBMSCs with PI3K inhibitor ( LY249002 ) on day 14 (n = 9). n. Expression of Alp and Runx2 after 7 days of osteogenic induction by qRT-PCR (n = 9). o. Expression of Spp1 and Bglap after 14 days of osteogenic induction measured by qRT-PCR (n = 9). OM: mBMSCs expressing vector after 7 days of osteogenesis induction; OM_IRF1: mBMSCs over-expressing Irf1 after 7 days of osteogenesis induction. Data are shown as the mean ± SD. *indicates a significant difference from the control group, * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The antibodies used in this study were listed below: GAPDH (HX1832; Huaxingbio, Beijing, China), PI3K kinase p85α (60225-1-Ig; Proteintech, Wuhan, China), AKT (10176-2-AP; Proteintech), p-PI3K P85α/P55γ/P85β-Y467/Y199/Y464 (AP0854; ABclonal, Wuhan, China), p-AKT1-S473 (AP0140; ABclonal), HRP-goat anti-mouse IgG (H + L) (HX2032; Huaxingbio) and HRP-goat anti-rabbit IgG (H + L) (ZB 2301; ZSGB-Bio, Beijing, China).

Techniques: Gene Expression, Comparison, Protein-Protein interactions, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Staining, Activity Assay, Over Expression, Quantitative RT-PCR, Plasmid Preparation, Control