Osteopontin Antibody - #AF0227

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. 1. Effect of kaempferol on renal CaOx crystal deposition and tubular injury in vivo. Eligible mice were divided into five groups (n = 8). After a period of 10-day drug treatments, all mice were sacrificed on the 11 th day for the following tests. Pathological results were shown by the (a) images as well as (b-g) corresponding quantitative analysis through HE staining, Pizzolato staining, PAS staining, TUNEL staining, OPN and CD44 immunohistochemistry staining of paraffin embedded kidney sections. (h-j) Renal dysfunction was determined by serum levels of BUN, creatinine and NAGL among the five groups. (k) Group annotations were applied for all histograms. Data were expressed as the fold changes of experimental group to NC group or GA group, and were represented as means ± SD. * p < 0.05 vs. NC group; ** p < 0.05 as GA+Kae (25mg/kg) vs. GA group, GA + Kae (50 mg/kg) vs. GA group, and GA + Kae (25 mg/kg) vs. GA + Kae (50 mg/kg) group, respectively.

FIGURE 8 The expression of osteogenesis-related genes and proteins: (A) Immunofluorescent images of Col-I and OPN expressed by BMSCs cultured on the different microcarriers for 14 days which were observed under CLSM (Scale bar = 50 μm); (B,C) qRT-PCR analysis of Col-I and OPN. (*p < 0.05, **p < 0.01, n = 3).

Figure 3 Hypoxia promotes proliferation, migration and odontoblastic differentiation in HDPCs. (A) Representative images of wound healing assays in the normoxic or hypoxic group. Scale bar, 1 mm. (B) Percentages of wound closure from three independent experiments are quantified. (C) Representative images of migratory cells stained with crystal violet. (D) Statistical quantification of the number of migratory cells from three independent experiments are shown. Scale bar, 100 µm. (E) Cell viability of HDPCs was detected using Cell Counting Kit-8 analysis. (F) ALP staining and (G) activity in the normoxic or hypoxic group on day 3. Scale bar, 200 µm. (H) Representative western blotting images showing the protein expression levels of RUNX2, Col I, OSX and OPN in the normoxic or hypoxic group on day 3, (I) which were quantified. Results are presented as the means ± SD from ≥ three independent experiments. **P<0.01 and ***P<0.001 vs. normoxia. Nor, normoxia; Hypo, hypoxia; OD, optical density; ALP, alkaline phosphatase; RUNX2, runt-related transcription factor 2; Col I, collagen type I; OSX, osterix; OPN, osteopontin.

Figure 4 The impact of C/EBPα in angiotensin II- (Ang II-) treated VSMCs. The relative protein expressions in Ang II-treated VSMCs with presence of C/EBPα overexpression or knockdown were analyzed by Western blot. GAPDH was used as an internal control. Data are presented as means ± SEM. (a, b) Evaluation of the influence of C/EBPα on the expression of α-SMA, SM-MHC, and OPN proteins. (c, d) Evaluation of the influence of C/EBPα on the expression of PIK3C2A protein. (e, f) Evaluation of the influence of C/EBPα on the expression of Beclin-1, p62, LC3II, and LC3I proteins. (g, h) Evaluation of the influence of C/EBPα on the expression of MMP-2 and MMP-9 proteins. (i) Intracellular ROS was measured by flow cytometry using DCFH-DA probe with presence of C/EBPα overexpression or knockdown. Control: control group; model: treated with 1 μM Ang II; vector: treated with 1 μM Ang II in vector transfected VSMCs; pcC/EBPα: treated with 1 μM Ang II in C/EBPα-overexpressed VSMCs; siNC: treated with 1 μM Ang II in siRNA NC-transfected VSMCs; siC/EBPα: treated with 1 μM Ang II in C/EBPα-knockdown VSMCs. ∗P < 0.05 vs. control,