PDLIM3 Antibody - #DF12525

Fig. 9 (A) Masson staining: the region between surrounding bone and scaffold (40x & 100x); (B) TRAP staining: TRAP positive cells (black arrows); (C) IHC staining of RUNX-2; (D) IHC staining of ALP; (E) Quantitative analysis for TRAP staining and IHC staining. * Compared with CPS group; # compared with Sr-5 group.

Figure 5. In vitro biocompatibilities and osteogenic activities of the substrates. (A) CCK-8 assay of hBMSCs on the indicated surfaces after 1, 3 and 7 days of culture. * denotes p < 0.05 and ** denotes p < 0.01 compared to Ti-NTs, # denotes p < 0.05 compared to the corresponding control group without peptides. Error bars denote the standard deviations over quadruplicate measurements with separately implants. (B) Confocal fluorescence microscopy images of hBMSCs stained with vinculin, F-actin and DAPI after being cultured for 24 h. Scale bar, 50 μm. (C-F) qRT-PCR assay of osteogenic gene expression of (C) ALP, (D) RUNX-2, (E) COL1 and (F) OPN of hBMSCs after 7 and 14 days of culture. * denotes p < 0.05, ** denotes p < 0.01 and *** denotes p < 0.001 compared to Ti-NTs-P-A; # denotes p < 0.05, & denotes p < 0.01 and $ denotes p < 0.001 compared to the corresponding control group without peptides. All error bars denote the standard deviations over quadruplicate measurements with separately implants. (G) Immunofluorescence staining of hBMSCs cultured on Ti-NTs-P-A for 7 days (ALP and RUNX-2) and 14 days (OPN). The images were obtained by confocal fluorescence microscopy. Scale bar, 50 μm. (H) Western blotting of hBMSCs cultured on the substrates for 7 and 14 days. At each time point, left lane was Ti-NTs, middle lane was Ti-NTs-A and right lane was Ti-NTs-P-A. * denotes p < 0.05, ** denotes p < 0.01 and *** denotes p < 0.001 compared to Ti-NTs-P-A. Error bars denote the standard deviations over triplicate measurements with separately Western blotting results.

Fig. 3 10− 6 M insulin promotes the osteogenic differentiation of human DPSCs. A 10− 6 M insulin up-regulated the mRNA levels of COL-1, ALP, OCN, and RUNX2 in DPSCs at day 3 and day 7 (n = 3). B 10− 6 M insulin promoted the protein expressions of COL-1, ALP, OCN, and RUNX2 in DPSCs at day 7 (n = 3). Representative western blotting (left) and quantification analysis (right). Full-length blots/gels are presented in Supplementary Fig. 1. C 10− 6 M insulin increased the secretion of extracellular bone metabolism and biochemical markers in DPSCs at day 1–7 and day 7–14 (n = 6). D 10− 6 M insulin enhanced alkaline phosphatase staining of DPSCs at day 21 (scale bars: 100 μm). E 10− 6 M insulin enhanced Alizarin red staining of DPSCs at day 21 (scale bars: 100 μm). F 10− 6 M insulin promoted the mineralized matrix formation of DPSCs at day 21 (n = 6). Data are expressed as the mean ± SD. *P 

Fig. 4 |Inactivation of ERK/MAPK signaling pathway inhibited the odontogenic differentiation of BMMSCs induced by TGC-CM. a ERK inhibitor U0126 significantly inhibited the mineralization and b reduced ALP, DSP, and DMP-1 protein levels of BMMSCs during odontogenic differentiation induced by TGC-CM for 7 days. Scale bar = 100 μm

Fig. 2. Comparison of osteogenic differentiation ability between NC and DOP(diabetic osteoporosis) hJBMMSCs. A. The ALP staining area was smaller and the color was lighter in DOP group than NC group on the 10th day (Mean ± SD, n = 6). B. The ALP activity was lower in DOP group than NC group after 7 days and 14 days of osteogenic induction (Mean ± SD, n = 6). C. After osteogenic induction, ARS (alizarin red S) staining on the 21st day showed that the number of mineralized nodules in DOP group was significantly less than that in NC group (Mean ± SD, n = 6). D. semiquantitative analysis showed that the OD (optical density) value of DOP group was significantly lower than that of NC group (Mean ± SD, n = 6). E. After osteogenic induction, the mRNA expressions of RUNX2, BMP2,ALP, OCN, SP7 and DLX5 in DOP group were significantly lower than those in NC group (Mean ± SD, n = 3). F. The protein expressions of RUNX2, BMP2, ALP and DLX5 in DOP group were significantly lower than those in NC group (Mean ± SD, n = 3). **P < 0.01,***P < 0.001.

Figure 3 Iron-overloaded MC3T3-E1 cells are more capable of osteogenic mineralization when treated with MT. (a) Alkaline phosphatase expression in MC3T3-E1 cells in each group was determined using an ALP staining. Images obtained from a gross scanner are shown on the left (scale bar: 1 mm), enlarged images are displayed in the middle (magnification: 100 and scale bar: 250 μm), and the quantitative results of the gross scanner images are displayed on the right. (b) ALP, Runx2, BMP2, and OCN mRNA expressions in MC3T3-E1 cells in each group were determined by RT-PCR. (c) Alizarin red S staining was used to identify each group's osteogenic mineralization of MC3T3-E1 cells. Images obtained from a gross scanner are shown on the left (scale bar: 1 mm), magnified images are displayed in the middle (magnification: 100 and scale bar: 250 μm), and a quantitative analysis of the left gross scanning images is displayed on the right. (d) The protein expression of OCN, ALP, Runx2, and BMP2 in MC3T3-E1 cells was determined by using Western blotting. ALP stands for alkaline phosphatase, RUNX2 for runt-related transcription factor 2, OCN for osteocalcin, and BMP2 for bone morphogenetic protein 2;