rabbit anti mouse runx2  (Cell Signaling Technology Inc)

Journal: Journal of Nanobiotechnology

Article Title: Apoptotic vesicles rescue impaired mesenchymal stem cells and their therapeutic capacity for osteoporosis by restoring miR-145a-5p deficiency

doi: 10.1186/s12951-024-02829-2

Figure Lengend Snippet: WT apoVs obviously rescued the impaired therapeutic capacity of OVX MSCs on the recovery of osteogenic differentiation, adipogenic differentiation and immunoregulatory properties of MSCs derived from the osteoporotic mice, rather than OVX apoVs. ( A , B ) BrdU labeling and continuous passage assay showed that MSCs from the OVX mice proliferated at a higher rate. Exogenous MSC infusion had no obvious effect on the proliferation of recipient MSCs from the OVX mice. ( C ) Alizarin red staining showed the recovery of osteogenic differentiation capacity of recipient MSCs from the OVX mice after exogenous infusion of WT MSCs and WT apoVs preconditioned OVX MSCs. ( D ) Western blot assay evaluating the expression of osteogenic differentiation-related protein Runx2 and ALP showed similar results as ( C ). ( E ) Oil red O staining manifested the recovery of adipogenic differentiation capacity of recipient MSCs from the OVX mice after exogenous infusion by the WT MSCs and WT apoVs preconditioned OVX MSCs. ( F ) Western blot assay evaluating the expression of adipogenic differentiation-related protein PPAR-γ and LPL showed similar results as ( E ). ( G , H ) Flow cytometric analysis showed that the WT apoVs preconditioned OVX MSCs infusion effectively rescued the immunoregulatory capacity of recipient MSCs, indicating inducing a similar amount of T lymphocytes apoptosis compared to the WT MSCs infusion. n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001. Data are presented as mean ± SD. Scale bar, 50 μm ( E )

Article Snippet: Anti-mouse cleaved caspase-3 (#9661), Anti-mouse Runx2 (#8486), Anti-mouse phospho-p44/42 MAPK (Erk1/2) (#4370), Anti-mouse p44/42 MAPK (Erk1/2) (#9102), Anti-mouse PI3 Kinase p110 (#4249), Anti-mouse PI3 Kinase p85 (#4292), Anti-mouse NF-κB p65 (#8242), Anti-mouse phospho-NF-κB p65 (#3033) antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA) and diluted to 1:1000 before using.

Techniques: Derivative Assay, Labeling, Staining, Western Blot, Expressing

Journal: Journal of Nanobiotechnology

Article Title: Apoptotic vesicles rescue impaired mesenchymal stem cells and their therapeutic capacity for osteoporosis by restoring miR-145a-5p deficiency

doi: 10.1186/s12951-024-02829-2

Figure Lengend Snippet: WT apoVs prominently remedy the impaired osteogenic differentiation, adipogenic differentiation and immunoregulatory properties of OVX MSCs in vitro, other than OVX apoVs. (A , B) BrdU labeling and continuous passage assay demonstrated that apoVs treatment had no obvious effect on the proliferation of OVX MSCs. (C) Alizarin red staining showed the recovery of osteogenic differentiation capacity of OVX MSCs after WT apoVs treatment, other than OVX apoVs. (D) Western blot assay indicated that the expression of osteogenic differentiation-related protein Runx2 and ALP was extremely elevated after WT apoVs treatment, instead of OVX apoVs treatment. (E) Oil red O staining manifested the recovery of adipogenic differentiation capacity of OVX MSCs stimulated by WT apoVs, rather than OVX apoVs. (F) Western blot assay indicated that the expression of adipogenic differentiation-related protein PPAR-γ and LPL was significantly reduced after WT apoVs treatment, but not OVX apoVs treatment. (G , H) Flow cytometric analysis demonstrated that WT apoVs treatment remarkably improved the immunoregulation capacity of OVX MSCs, while the OVX apoVs treated group showed no similar effect. n = 3. * p < 0.05, ** p < 0.01. Data are presented as mean ± SD. Scale bar, 50 μm (E)

Article Snippet: Anti-mouse cleaved caspase-3 (#9661), Anti-mouse Runx2 (#8486), Anti-mouse phospho-p44/42 MAPK (Erk1/2) (#4370), Anti-mouse p44/42 MAPK (Erk1/2) (#9102), Anti-mouse PI3 Kinase p110 (#4249), Anti-mouse PI3 Kinase p85 (#4292), Anti-mouse NF-κB p65 (#8242), Anti-mouse phospho-NF-κB p65 (#3033) antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA) and diluted to 1:1000 before using.

Techniques: In Vitro, Labeling, Staining, Western Blot, Expressing

Journal: Journal of Nanobiotechnology

Article Title: Apoptotic vesicles rescue impaired mesenchymal stem cells and their therapeutic capacity for osteoporosis by restoring miR-145a-5p deficiency

doi: 10.1186/s12951-024-02829-2

Figure Lengend Snippet: miR-145a-5p contributed to the WT apoVs-mediated rescue of impaired OVX MSCs. ( A ) Real-time PCR analysis indicated that miR-145a-5p expression was decreased in OVX MSCs as compared with WT MSCs, which was increased when treated with WT apoVs in vitro. ( B ) Real-time PCR analysis indicated that miR-145a-5p expression was decreased in OVX apoVs compared to WT apoVs. ( C ) Real-time PCR analysis showed that pri-miR-145a-5p expression was downregulated in OVX MSCs compared to WT MSCs, which was unchanged after apoVs stimulation. ( D ) Real-time PCR analysis indicated that actinomycin D treatment failed to affect the miR-145a-5p expression in WT apoVs treated OVX MSCs. ( E , F ) Western blot assay indicated that miR-145a-5p mimics downregulated TGF-β1/Smad2/3 signaling pathway and upregulated Wnt/β-catenin signaling pathway in OVX MSCs, while miR-145a-5p inhibitor displayed the opposite effect in WT MSCs. ( G , H ) Alizarin red staining showed that miR-145a-5p mimics promoted the mineralized nodule formation in OVX MSCs, while miR-145a-5p inhibitor displayed the opposite effect in WT MSCs. ( I , J ) Western blot assay indicated that miR-145a-5p mimics promoted the expression of osteogenic differentiation-related protein Runx2 and ALP in OVX MSCs, while miR-145a-5p inhibitor displayed the opposite effect in WT MSCs. ( K , L ) Flow cytometric analysis indicated that miR-145a-5p mimics upregulated the immunoregulatory capacity of OVX MSCs, while miR-145a-5p inhibitor displayed the opposite effect in WT MSCs. n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001. Data are presented as mean ± SD

Article Snippet: Anti-mouse cleaved caspase-3 (#9661), Anti-mouse Runx2 (#8486), Anti-mouse phospho-p44/42 MAPK (Erk1/2) (#4370), Anti-mouse p44/42 MAPK (Erk1/2) (#9102), Anti-mouse PI3 Kinase p110 (#4249), Anti-mouse PI3 Kinase p85 (#4292), Anti-mouse NF-κB p65 (#8242), Anti-mouse phospho-NF-κB p65 (#3033) antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA) and diluted to 1:1000 before using.

Techniques: Real-time Polymerase Chain Reaction, Expressing, In Vitro, Western Blot, Staining