anti ocn primary antibody  (Proteintech)

Journal: World Journal of Stem Cells

Article Title: EZH2, via an association with KDM2B, modulates osteogenic differentiation of root apical papillary stem cells

doi: 10.4252/wjsc.v17.i4.103482

Figure Lengend Snippet: Knockdown of enhancer of zeste homolog 2 inhibits steo/dentinogenic differentiation potential of human apical papillary stem cells. A: Quantitative polymerase chain reaction showed that the expression of enhancer of zeste homolog 2 (EZH2) was inhibited in human apical papillary stem cells (hSCAPs); B: Western blot analysis confirmed the knockdown of EZH2 in hSCAPs; C: Knockdown of EZH2 decreased alkaline phosphatase activity in hSCAPs; D and E: Alizarin red staining and quantitative calcium analysis demonstrated that knockdown of EZH2 inhibited mineralization in hSCAPs; F-H: Quantitative polymerase chain reaction showed that knockdown of EZH2 downregulated mRNA expression levels of bone sialoprotein (F), dentin sialophosphoprotein (G), and osteocalcin (H) in hSCAPs. GAPDH and ACTB was used as the internal controls. Data are presented as the mean ± SD ( n = 3). Statistical analysis was performed using Student’s t -test. a P ≤ 0.05, b P ≤ 0.01, c P ≤ 0.001. EZH2: Enhancer of zeste homolog 2; BSP: Bone sialoprotein; DSPP: Dentin sialophosphoprotein; OCN: Osteocalcin.

Article Snippet: Immunohistochemical analysis followed established protocols[ ] using primary antibodies against osteocalcin (OCN) (Bioss bs-4917R) and DSPP (Bioss bs10316R).

Techniques: Knockdown, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Activity Assay, Staining

Journal: World Journal of Stem Cells

Article Title: EZH2, via an association with KDM2B, modulates osteogenic differentiation of root apical papillary stem cells

doi: 10.4252/wjsc.v17.i4.103482

Figure Lengend Snippet: Overexpression of enhancer of zeste homolog 2 enhances osteo/dentinogenic differentiation potential of human apical papillary stem cells. A: Quantitative polymerase chain reaction showed that enhancer of zeste homolog 2 (EZH2) was overexpressed in human apical papillary stem cells (hSCAPs); B: Western blot analysis confirmed overexpression of EZH2 in hSCAPs; C: Overexpression of EZH2 increased alkaline phosphatase activity in hSCAPs; D and E: Alizarin red staining and quantitative calcium analysis results demonstrated that overexpression of EZH2 enhanced mineralization in hSCAPs; F-H: Quantitative polymerase chain reaction showed that overexpression of EZH2 upregulated mRNA expression levels of bone sialoprotein (F), dentin sialophosphoprotein (G), and osteocalcin (H) in hSCAPs; I: Hematoxylin-eosin staining and quantitative measurement showed that overexpression of EZH2 promoted bone/dentin-like tissue formation. Scale bar = 100 μm (B: Bone/dentin-like tissues; HA: Hydroxyapatite tricalcium carrier; CT: Connective tissue); J: Immunohistochemical staining and quantitative analysis of dentin sialophosphoprotein and bone sialoprotein. GAPDH and ACTB were used as the internal controls. Data are presented as the mean ± SD ( n = 3). Statistical analysis was performed using Student’s t -test. a P ≤ 0.05, b P ≤ 0.01, c P ≤ 0.001. EZH2: Enhancer of zeste homolog 2; BSP: Bone sialoprotein; DSPP: Dentin sialophosphoprotein; OCN: Osteocalcin.

Article Snippet: Immunohistochemical analysis followed established protocols[ ] using primary antibodies against osteocalcin (OCN) (Bioss bs-4917R) and DSPP (Bioss bs10316R).

Techniques: Over Expression, Real-time Polymerase Chain Reaction, Western Blot, Activity Assay, Staining, Expressing, Immunohistochemical staining

Journal: Nature Communications

Article Title: High-throughput bioprinting of spheroids for scalable tissue fabrication

doi: 10.1038/s41467-024-54504-7

Figure Lengend Snippet: A HITS-Bio setup under surgical settings. Inset images (1–2) demonstrate deposited BONink and spheroid placement, (ii) created calvarial defects ( ~ 5 mm in a diameter), and (iii) bioprinted bone constructs with BONink and spheroids. Scale bar: 5 mm. B Visualization of newly regenerated bone in calvarial defects with transverse and sagittal planes at Weeks 3 and 6 via µCT, including empty, BONink only, low-density and high-density groups. Scale bar: 1 mm. C Relevant quantification for bone regeneration within the defect examined at Weeks 3 and 6 ( n = 7 independent defects, one-way ANOVA, all other shown comparisons p < 0.0001), including (i) BV/TV (new bone volume to total bone volume, %) (Week 3: a vs. b p = 0.35139, a vs. c p = 0.00074, a vs. d p = 0.00545, b vs. c p = 0.04183, b vs. d p = 0.20350, c vs. d p = 0.85065, Week 6: a vs. b p = 0.60072, a vs. c p = 0.00186, a vs. d p = 0.00212, b vs. c p = 0.03614, b vs. d p = 0.04048, c vs. d p = 0.99995), (ii) normalized BMD (bone mineral density, %) (Week 3: a vs. b p = 0.045, b vs. c p = 0.03277, b vs. d p = 0.07177, c vs. d p = 0.98269, Week 6: a vs. b p = 0.63421, a vs. c p = 0.00473, a vs. d p = 0.00528, b vs. c p = 0.07204, b vs. d p = 0.07910, c vs. d p = 0.99996), (iii) bone coverage area (%) (Week 3: a vs. b p = 0.06611, b vs. c p = 0.00656, b vs. d p = 0.00464, c vs. d p = 0.99893, Week 6: a vs. b p = 0.11033, b vs. c p = 0.00259, b vs. d p = 0.00032, c vs. d p = 0.83536) (The outlier was not included in the data plot), and (iv) scores for bony bridging (Week 3: a vs. b p = 0.03791, b vs. c p = 0.01013, b vs. d p = 0.01013, c vs. d p = 1.0, Week 6: a vs. b p = 0.04481, b vs. c p = 0.00326, b vs. d p = 0.00326, c vs. d p = 1.0). D Mechanical properties of the retrieved defect area 6 weeks after the surgery ( n = 3, 4, 4, 3 independent defects (from left to right), one-way ANOVA) (Shear yield strength: a vs. b p = 0.97205, a vs. c p = 0.95577, a vs. d p = 0.14142, b vs. c p = 0.74719, b vs. d p = 0.05602, c vs. d p = 0.23294) (Modulus of resilience: a vs. b p = 0.98816, a vs. c p = 0.99588, a vs. d p = 0.00672, b vs. c p = 0.99957, b vs. d p = 0.00692, c vs. d p = 0.00598) (Shear modulus: a vs. b p = 0.65248, a vs. c p = 0.95136, a vs. d p = 0.94709, b vs. c p = 0.31149, b vs. d p = 0.92926, c vs. d p = 0.69672). E Histomorphometric characterization of sectioned defect after decalcification and stained for H&E (scale bar: 500 µm), MT (scale bar: 1 mm), and IHC (P1NP and OCN) (scale bar: 500 µm). Representative images were obtained from at least three independent repetitions. Data are presented as mean ± SD where * p < 0.05, ** p < 0.01, and *** p < 0.001. Source data are provided as a Source Data file.

Article Snippet: To detect the bone tissue, sections were incubated with a mouse anti-procollagen 1 N-Peptide (P1NP) primary antibody (1:250 in 2.5% NGS, MA5-51183, Invitrogen), mouse anti- osteocalcin (OCN) primary antibody (1:200 in 2.5% NGS, 33-5400, Invitrogen), mouse anti-RUNX2 (runt-related transcription factor 2) primary antibody (1:20 in 2.5% NGS, ab76956; Abcam) and a rabbit anti-Sp7/Osterix (OSTERIX) primary antibody (1:100 in 2.5% NGS; ab209484, Abcam) or a rabbit anti-bone sialoprotein (BSP) primary antibody (1:100 in 2.5% NGS; ab52128, Abcam) overnight.

Techniques: Construct, Shear, Staining