Fig 1: Runx2 is a potential target of miR-488. (A) Construction profile of the pLUC-Runx2 vector is presented in the diagram, which contained the miR-488 target sites in Runx2 3'UTR-wt, red sequence presented the mutation region. Reverse transcription-quantitative PCR showed that (B) miR-488 mimics and (C) miR-488 inhibitor were successfully transfected into cells. (D) Reverse transcription-quantitative PCR showed cotransfection of miR-488 mimics and miR-488 inhibitor. Reverse transcription-quantitative PCR showed that (E) pcDNA 3.1-Runx2 was successfully transfected into cells. *P<0.05. (F) Relative luciferase activity demonstrated that Runx2 is a potential target of miR-488. *P<0.05 vs. the miR control group; #P<0.05 vs. the miR-488 mimics group. Runx2, runt-related transcription factor 2; miR, microRNA; 3'UTR, 3'-untranslated region; wt, wild-type; mut, mutant; ns, not significant.
Fig 2: Assessing the expressions of differentiation-related regulatory genes in the long bone of mouse embryos from control and PGDM group. A-C: Representative images of Sox9 (A), Osterix (B) and Runx2 (C) immunofluorescence in the growth plate and diaphysis of E18.5 mouse embryos from control and PGDM group. Cell nuclei were counterstained with DAPI. Right panels are the high magnification images indicated by dotted squares in the middle panels. D-F: Bar charts showing the ratio comparisons of Sox9+ (D), Runx2+ (E) and Osterix+ (F) cell numbers of E18.5 mouse embryos from control and PGDM groups. G-I: Quantitative PCR data showing the mRNA expression of Sox9 (G), Runx2 (H) and Osterix (I) from E18.5 control and PGDM mouse embryos. J-L: Western blot data showing the expression of Sox9 (J), Runx2 (K) and Osterix (L) from E18.5 control and PGDM mouse embryos. Scale bars = 400 μm in the left and middle panels of A-C, 50 μm in the right panels of A-C. For D, n = 9, for E, n = 7, for F, n = 6, for G-I, n = 3. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.
Fig 3: Proposed model of psoralen-induced osteogenic proliferation and differentiation through the TGF-ß/Smad3 pathway in human bone marrow mesenchymal stem cells. BMP4, bone morphogenetic protein 4; OPN, osteopontin; Runx2, runt-related transcription factor 2; TßR, TGF-ß receptor; p, phosphorylated.
Fig 4: ER stress plays an important role in bone formation and is regulated by salubrinal (SAL) concentration. A-B, Osteogenic induction in POBs for 7 d with salubrinal administered at 0.2, 1 or 5 µmol/L in the presence of ER stress inducer tunicamycin (Tm; 100 ng/mL). After 48 h, ALP expression (black, 100×) decreased with Tm treatment, which was reversed by SAL. C, CCK8 assay for POBs showing the same trend. D-E, Representative Western blots showing SAL’s protection of osteogenesis against ER stress. The expression of Runx2/ß-actin protein increased and the expression of p-eIF2a/eIF2a decreased with recovery of the ER. Data were presented as the ratio with ß-actin and mean ± SD. The asterisks (*) represent P < .05. VC, vehicle control
Fig 5: ER stress occurred in alveolar bone after tooth extraction. A, Representative Western blots of the protein expression levels of ER stress-related proteins eIF2a, p-eIF2a and osteogenic marker Runx2 in bone tissue at tooth extraction sites 7 and 14 d after tooth extraction. B, p-eIF2a/eIF2a and Runx2/ß-actin proteins was expressed on days 7 and 14. C-D, Representative photomicrographs of the stained sections of extraction sockets. The blue dotted line indicates the tooth extraction socket, the star indicates irregular woven bone, and the blue arrow indicates the bone bridge. 14 d significantly increased the number of collagen fibres. Top, HE staining, 50×; bottom, Masson trichrome staining, left 50×, right 200×. Blue-stained fibres in the connective tissue indicate collagen. Data were presented as the ratio with ß-actin and mean ± SD. The asterisk (*) represents P < .05 (n = 12)
Supplier Page from Abcam for Anti-RUNX2 antibody