recombinant human cdk9 cyclin t1 proteins c40 18cg  (Sino Biological)

Journal: Advanced Science

Article Title: Steap4 Promotes Senile Osteoporosis via Fe 2+ ‐ROS/C/EBPβ Feedback‐Driven Ferroptosis and Adipogenesis in Senescent BMSCs

doi: 10.1002/advs.202509926

Figure Lengend Snippet: Ferroptosis of osteoblasts is involved in osteoporosis during ageing. A) Femoral micro‐CT scanning and 3D reconstruction. B) Bone volume/total volume (BV/TV) and C) trabecular separation (Tb.Sp.) were analyzed using femoral 3D reconstruction data ( n = 7 per group). D) H&E staining (scale bar: 100 µm). E) Immunohistochemistry (IHC) staining for p16 (scale bar: 100 µm). F) Quantitative analysis of p16(+) cells ( n = 4 per group). G,H) KEGG enrichment analysis. I) Prussian blue staining: the blue particles are iron‐containing particles (scale bar: 50 µm). J) BODIPY 581/591 C11 was used to detect lipid peroxides. K) DCFH‐DA was used to detect ROS, and images were observed using fluorescence microscopy and statistically quantified by flow cytometry (scale bar: 50 µm). L) Detection of intracellular Fe 2+ . M) JC‐1 was used to evaluate mitochondrial membrane potential, observed via fluorescence microscopy and quantified via flow cytometry (scale bar: 50 µm). (The data are presented as the means ± SDs. Statistical analysis was performed using one‐way ANOVA. *** p < 0.001, ** p < 0.01, * p < 0.05, in vitro experiments n = 4.).

Article Snippet: Tissue cyclin‐dependent kinase inhibitor 2A (p16) (Proteintech, WuHan, #10883‐1‐AP), PPARG (Proteintech, WuHan, #16643‐1‐AP), OCN (ABclonal, WuHan, #A6205), Steap4 (Proteintech, WuHan, #11944‐1‐AP), solute carrier family 11 (proton divalent metal ion transporters) member 2 (Slc11a2 or DMT1) (Proteintech, WuHan, #20507‐1‐AP), and C/EBPβ (ABclonal, WuHan, #A0711) were detected.

Techniques: Micro-CT, Staining, Immunohistochemistry, Fluorescence, Microscopy, Flow Cytometry, Membrane, In Vitro

Journal: Advanced Science

Article Title: Steap4 Promotes Senile Osteoporosis via Fe 2+ ‐ROS/C/EBPβ Feedback‐Driven Ferroptosis and Adipogenesis in Senescent BMSCs

doi: 10.1002/advs.202509926

Figure Lengend Snippet: BMSCs from ageing mice are more susceptible to ferroptosis. A) Senescence β‐Gal staining (scale bar: 50 µm), immunofluorescence staining of p16 (red for p16 and blue for nuclei, scale bar: 20 µm) and p21 (red for p21 and blue for nuclei, scale bar: 20 µm). B) ALP staining, ARS staining and FerroGreen Assay (green for FerroGreen and blue for Hoechst, scale bar: 20 µm). C–E) Quantitative analysis of β‐Gal, p16, and p21. F–H) Quantitative analysis of ALP staining, ARS staining and FerroGreen Assay. I) Images of the BMSCs after coimmunostaining: red (no‐ox‐Bodipy for non‐lipid peroxidation), green (ox‐Bodipy for lipid peroxidation) and blue (hoechst, scale bar: 20 µm). J) GPX4, FTL, and FTH1 protein levels, detected by western blot analysis. Quantitative analysis of K) ox‐Bodipy, L) GPX4, M) FTL, and N) FTH1. O) Senescence β‐Gal staining, ALP staining and ARS staining (scale bar: 50 µm). Quantitative analysis of P) β‐Gal staining, Q) ALP staining, and R) ARS staining. (The data are presented as the means ± SDs. Statistical analysis was performed using one‐way ANOVA. *** p < 0.001, ** p < 0.01, * p < 0.05, ns: not significant, n = 4.).

Article Snippet: Tissue cyclin‐dependent kinase inhibitor 2A (p16) (Proteintech, WuHan, #10883‐1‐AP), PPARG (Proteintech, WuHan, #16643‐1‐AP), OCN (ABclonal, WuHan, #A6205), Steap4 (Proteintech, WuHan, #11944‐1‐AP), solute carrier family 11 (proton divalent metal ion transporters) member 2 (Slc11a2 or DMT1) (Proteintech, WuHan, #20507‐1‐AP), and C/EBPβ (ABclonal, WuHan, #A0711) were detected.

Techniques: Staining, Immunofluorescence, Western Blot

Journal: Advanced Science

Article Title: Steap4 Promotes Senile Osteoporosis via Fe 2+ ‐ROS/C/EBPβ Feedback‐Driven Ferroptosis and Adipogenesis in Senescent BMSCs

doi: 10.1002/advs.202509926

Figure Lengend Snippet: Steap4 is involved in senescent BMSCs ferroptosis. A) Venn analysis. B) The top 10 DEGs identified by Venn diagram analysis. C–E) Steap4 expression in BMSCs from 4‐month‐old and 16‐month‐old mice ( n = 3 per group); and Steap4 expression was detected after extracting BMSCs from 4‐month‐old mice and culturing them into the first generation (P1) and fourth generation (P4). F) Detection of intracellular Fe 2+ . G) BODIPY 581/591 C11 was used to detect lipid peroxides. H) JC‐1 was used to evaluate the mitochondrial membrane potential. I) GPX4, FTL, and FTH1 protein levels, detected by western blot analysis. Quantitative analysis of J) GPX4, K) FTL, and L) FTH1. M) Senescence β‐Gal staining (scale bar: 50 µm), immunofluorescence staining of p16 (red for p16 and blue for nuclei, scale bar: 20 µm) and p21 (red for p21 and blue for nuclei, scale bar: 20 µm). Quantitative analysis of N) β‐Gal staining, O) p16, and P) p21. Q) ALP staining and ARS staining. R) Runx2, Osterix and OCN protein levels, detected by western blot analysis. Quantitative analysis of S) ALP staining, T) ARS staining, U) Runx2, V) Osterix, and W) OCN. (The data are presented as the means ± SDs. Statistical analysis was performed using one‐way ANOVA. *** p < 0.001, ** p < 0.01, * p < 0.05, ns: not significant, n = 4.).

Article Snippet: Tissue cyclin‐dependent kinase inhibitor 2A (p16) (Proteintech, WuHan, #10883‐1‐AP), PPARG (Proteintech, WuHan, #16643‐1‐AP), OCN (ABclonal, WuHan, #A6205), Steap4 (Proteintech, WuHan, #11944‐1‐AP), solute carrier family 11 (proton divalent metal ion transporters) member 2 (Slc11a2 or DMT1) (Proteintech, WuHan, #20507‐1‐AP), and C/EBPβ (ABclonal, WuHan, #A0711) were detected.

Techniques: Expressing, Membrane, Western Blot, Staining, Immunofluorescence

Journal: Advanced Science

Article Title: Steap4 Promotes Senile Osteoporosis via Fe 2+ ‐ROS/C/EBPβ Feedback‐Driven Ferroptosis and Adipogenesis in Senescent BMSCs

doi: 10.1002/advs.202509926

Figure Lengend Snippet: Steap4 promotes ferroptosis through the accumulation of Fe 2+ in BMSCs. A) Graphical summary of the hypothesis. B) Senescence β‐Gal staining, ALP staining and ARS staining (scale bar: 50 µm). Quantitative analysis of C) β‐Gal staining, D) ALP staining and E) ARS staining. F) FerroGreen Assay (scale bar: 20 µm). G) Quantitative analysis of FerroGreen. H) Detection of intracellular Fe 2+ . I) JC‐1 was used to evaluate the mitochondrial membrane potential. J) BODIPY 581/591 C11 was used to detect lipid peroxides. K) DMT1, GPX4, FTL, and FTH1 protein levels, detected by western blot analysis. Quantitative analysis of L) DMT1, M) GPX4, N) FTL, and O) FTH1. P) Graphical abstract of the animal models. Q) Micro‐CT scanning and 3D reconstruction. R) H&E staining (scale bar: 200 µm of the overall picture and 500 µm of the partial enlarged picture) and Masson staining: blue‐stained tissue indicates newly formed bone, and red indicates mature bone tissue (scale bar: 200 µm of the overall picture and 500 µm of the partial enlarged picture). S) Steap4 and DMT1 protein levels, detected by western blot analysis. Quantitative analysis of T) Steap4 and U) DMT1. V) IHC staining of p16 (scale bar: 50 µm). W) Quantitative analysis of p16. (The data are presented as the means ± SDs. Statistical analysis was performed using one‐way ANOVA. *** p < 0.001, ** p < 0.01, * p < 0.05, in vitro experiments n = 4, in vivo experiments n = 7.).

Article Snippet: Tissue cyclin‐dependent kinase inhibitor 2A (p16) (Proteintech, WuHan, #10883‐1‐AP), PPARG (Proteintech, WuHan, #16643‐1‐AP), OCN (ABclonal, WuHan, #A6205), Steap4 (Proteintech, WuHan, #11944‐1‐AP), solute carrier family 11 (proton divalent metal ion transporters) member 2 (Slc11a2 or DMT1) (Proteintech, WuHan, #20507‐1‐AP), and C/EBPβ (ABclonal, WuHan, #A0711) were detected.

Techniques: Staining, Membrane, Western Blot, Micro-CT, Immunohistochemistry, In Vitro, In Vivo

Journal: Advanced Science

Article Title: Steap4 Promotes Senile Osteoporosis via Fe 2+ ‐ROS/C/EBPβ Feedback‐Driven Ferroptosis and Adipogenesis in Senescent BMSCs

doi: 10.1002/advs.202509926

Figure Lengend Snippet: The Steap4/Fe 2+ ‐ROS/C/EBPβ/Steap4 axis is involved in senescent BMSCs ferroptosis. A) Venn analysis of transcription factors and 16‐month versus 4‐month RNA‐seq results. B) FPKM of Cebpa, Cebpb, Hes1 and C‐Jun in 16‐month versus 4‐month RNA‐seq data. C) Graphical summary of the hypothesis. D) FPKM of Cebpa, Cebpb, Hes1, and C‐Jun in 16‐month VS 16‐month+DFO RNA‐seq results. E) Western blot analysis of C/EBPβ. F) RT‒qPCR of Cebpb , Steap4 , and Dmt1 . G) Images of the BMSCs after coimmunostaining: red (no‐ox‐Bodipy for non‐lipid peroxidation), green (ox‐Bodipy for lipid peroxidation) and blue (hoechst, scale bar: 20 µm). H) Quantitative analysis of ox‐Bodipy. I) Steap4, DMT1, FTL, and FTH1 protein levels, detected by western blot analysis. Quantitative analysis of J) Steap4, K) DMT1, L) FTL, and M) FTH1. N) Images of the BMSCs after coimmunostaining: red (no‐ox‐Bodipy for non‐lipid peroxidation), green (ox‐Bodipy for lipid peroxidation) and blue (hoechst, scale bar: 20 µm). O) Immunofluorescence staining of p16 (red for p16 and blue for nuclei, scale bar: 20 µm) and p21 (red for p21 and blue for nuclei, scale bar: 20 µm). P) Steap4, DMT1 and FTH1 protein levels, detected by western blot analysis. Quantitative analysis of Q) ox‐Bodipy, R) p16, S) p21, T) Steap4, U) DMT1, and V) FTH1. W) ChIP‐qPCR: Schematic diagram of three ChIP‐qPCR sites in the promoter region of the Steap4 gene, and the ChIP‐qPCR results. (The data are presented as the means ± SDs. Statistical analysis was performed using one‐way ANOVA. *** p < 0.001, ** p < 0.01, * p < 0.05, n = 4.).

Article Snippet: Tissue cyclin‐dependent kinase inhibitor 2A (p16) (Proteintech, WuHan, #10883‐1‐AP), PPARG (Proteintech, WuHan, #16643‐1‐AP), OCN (ABclonal, WuHan, #A6205), Steap4 (Proteintech, WuHan, #11944‐1‐AP), solute carrier family 11 (proton divalent metal ion transporters) member 2 (Slc11a2 or DMT1) (Proteintech, WuHan, #20507‐1‐AP), and C/EBPβ (ABclonal, WuHan, #A0711) were detected.

Techniques: RNA Sequencing, Western Blot, Immunofluorescence, Staining, ChIP-qPCR