Journal: BMC Molecular and Cell Biology

Article Title: Compression and tension variably alter Osteoprotegerin expression via miR-3198 in periodontal ligament cells

doi: 10.1186/s12860-019-0187-2

Figure Lengend Snippet: miR-3198 gain-of-function and loss-of-function experiments. Results of real-time RT-PCR analysis for miR-3198 ( a , b ), OPG and RANKL ( c , d ) expression in HPL cells after transfection of miR-3198 inhibitor ( a , c ) and miR-3198 mimic ( b , d ). n = 3. Biological triplicated. Fold change from the control is shown. Open bar indicates the fold change of OPG expression, and close bar indicates that of RANKL expression ( c , d ). The concentrations of OPG as measured by ELISA after transfection of miR-3198 inhibitor ( e ) and miR-3198 mimic ( f ) are shown ( n = 3). Results of real-time PCR analysis for miR-1207 ( g , h ), OPG and RANKL ( i , j ) expression in HPL cells under the transfection of miR-1207 inhibitor ( g , i ) and miR-1207 mimic ( h , j ). n = 3. Biological triplicated. Fold changes from the control are shown. * indicates P < 0.05 versus control, and NS indicating there was no significant difference between samples

Article Snippet: The concentration of OPG in the culture supernatant was measured using an OPG ELISA kit (Boster Biological Technology, Pleasanton, CA), according to the manufacturer’s instructions.

Techniques: Quantitative RT-PCR, Expressing, Transfection, Control, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction

Journal: BMC Molecular and Cell Biology

Article Title: Compression and tension variably alter Osteoprotegerin expression via miR-3198 in periodontal ligament cells

doi: 10.1186/s12860-019-0187-2

Figure Lengend Snippet: miR-3198 regulates the mechanical stress-mediated change of OPG expression. Results of real-time RT-PCR analysis for OPG and RANKL expression in HPL cells in the compression ( a ) and tension ( b ) experiments. n = 3. Biological triplicated. Fold change from the control are shown. Cont, control; Inh, transfection of miR-3198 inhibitor; Mimic, transfection of miR-3198 mimic; press, compression; tens, tension; TF, transfection. Also shown are the OPG concentrations measured by ELISA in the compression ( c ) and tension ( d ) experiments ( n = 3). * indicates P < 0.05 versus control. † indicates P < 0.05 between samples. NS indicates there was no significant difference between samples. e and f Representative image of the western blotting for OPG was shown. g and h Relative band intensity of the western blotting for OPG. *: P < 0.05 versus control. †: P < 0.05 between the groups. NS, no significant difference between the samples

Article Snippet: The concentration of OPG in the culture supernatant was measured using an OPG ELISA kit (Boster Biological Technology, Pleasanton, CA), according to the manufacturer’s instructions.

Techniques: Expressing, Quantitative RT-PCR, Control, Transfection, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: International Journal of Medical Sciences

Article Title: HK2-mediated Glycolysis Inhibits Mineralization of Cementoblasts Under Compression by Suppressing the Piezo1/Wnt Signaling

doi: 10.7150/ijms.109287

Figure Lengend Snippet: Compression inhibited cementoblasts' mineralization and Piezo1 expression. (A) The relative mRNA levels of Piezo1 were detected by qRT-PCR after cementoblasts were subjected to 2.0 g/cm² compression for 12 hours under distinct buffer membranes. (B) and (C) The relative mRNA and protein levels of Piezo1, RANKL, OPG, RANKL/OPG, Runx2, OSX, and OPN were identified by qRT-PCR and WB. The blot images represented data from 3 independent experiments. The data are presented as mean ± SD (*P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001).

Article Snippet: The PVDF membranes were blocked at room temperature with 5% bovine serum albumin (BSA) for 1 hour, followed by incubation overnight at 4°C with primary antibodies against Piezo1 (1:1000, ABclonal, China), GLUT1 (1:5000, ABclonal, China), HK2 (1:1000, Abcam, UK), PFKFB3 (1:2000, Abcam, UK), LDHA (1:2000, Abcam, UK), β-catenin (1:1000, ABclonal, China), TCF1 (1:1000, ABclonal, China), LEF1 (1:1000, ABclonal, China), RANKL (1:1000, Proteintech, USA), OPG (1:1000, Proteintech, USA), Runx2 (1:1000, Cell Signaling Technology, USA), OSX (1:1000, Cell Signaling Technology, USA), and OPN (1:1500, Cell Signaling Technology, USA), all from rabbit sources.

Techniques: Expressing, Quantitative RT-PCR

Journal: International Journal of Medical Sciences

Article Title: HK2-mediated Glycolysis Inhibits Mineralization of Cementoblasts Under Compression by Suppressing the Piezo1/Wnt Signaling

doi: 10.7150/ijms.109287

Figure Lengend Snippet: Piezo1 mediated mechanotransduction in cementoblasts and promoted mineralization. (A) The knockdown efficiency of the three Piezo1 siRNAs was confirmed using qRT-PCR and WB. (B) The impact of varying doses of Yoda1 on cell viability after 24h was evaluated by the CCK-8 assay. (C) Intracellular calcium concentration was detected by the Fluo-4 AM calcium ion fluorescence probe. Images were captured by fluorescence microscopy. Scale bar = 50μm. (D) and (E) The relative mRNA and protein levels of RANKL, OPG, RANKL/OPG, Runx2, OSX, and OPN were identified using qRT-PCR and WB. The blot images represented data from three independent experiments. (F) Von Kossa (VK), Alizarin Red S (ARS) (after 3 weeks of mineralization induction) and alkaline phosphatase staining (after 2 weeks of mineralization induction). Scale bar = 10μm. The data are presented as mean ± SD (*P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001). Con: Control, Pz1: Piezo1, F: Force.

Article Snippet: The PVDF membranes were blocked at room temperature with 5% bovine serum albumin (BSA) for 1 hour, followed by incubation overnight at 4°C with primary antibodies against Piezo1 (1:1000, ABclonal, China), GLUT1 (1:5000, ABclonal, China), HK2 (1:1000, Abcam, UK), PFKFB3 (1:2000, Abcam, UK), LDHA (1:2000, Abcam, UK), β-catenin (1:1000, ABclonal, China), TCF1 (1:1000, ABclonal, China), LEF1 (1:1000, ABclonal, China), RANKL (1:1000, Proteintech, USA), OPG (1:1000, Proteintech, USA), Runx2 (1:1000, Cell Signaling Technology, USA), OSX (1:1000, Cell Signaling Technology, USA), and OPN (1:1500, Cell Signaling Technology, USA), all from rabbit sources.

Techniques: Knockdown, Quantitative RT-PCR, CCK-8 Assay, Concentration Assay, Fluorescence, Microscopy, Staining, Control

Journal: International Journal of Medical Sciences

Article Title: HK2-mediated Glycolysis Inhibits Mineralization of Cementoblasts Under Compression by Suppressing the Piezo1/Wnt Signaling

doi: 10.7150/ijms.109287

Figure Lengend Snippet: The Wnt/β-catenin signaling promoted mineralization of cementoblasts. (A) The effects of different concentrations of DKK1 and LiCl on cell viability after 24 hours was evaluated using the CCK-8 assay. (B) and (C) The relative mRNA and protein levels of β-catenin and its target transcription factors TCF1 and LEF1 were detected by qRT-PCR and WB. (D) and (E) The relative mRNA and protein levels of RANKL, OPG, RANKL/OPG, Runx2, OSX, and OPN were identified by qRT-PCR and WB. The blot images represented data from 3 independent experiments. The data are presented as mean ± SD (*P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001). Con: Control, F: Force.

Article Snippet: The PVDF membranes were blocked at room temperature with 5% bovine serum albumin (BSA) for 1 hour, followed by incubation overnight at 4°C with primary antibodies against Piezo1 (1:1000, ABclonal, China), GLUT1 (1:5000, ABclonal, China), HK2 (1:1000, Abcam, UK), PFKFB3 (1:2000, Abcam, UK), LDHA (1:2000, Abcam, UK), β-catenin (1:1000, ABclonal, China), TCF1 (1:1000, ABclonal, China), LEF1 (1:1000, ABclonal, China), RANKL (1:1000, Proteintech, USA), OPG (1:1000, Proteintech, USA), Runx2 (1:1000, Cell Signaling Technology, USA), OSX (1:1000, Cell Signaling Technology, USA), and OPN (1:1500, Cell Signaling Technology, USA), all from rabbit sources.

Techniques: CCK-8 Assay, Quantitative RT-PCR, Control

Journal: International Journal of Medical Sciences

Article Title: HK2-mediated Glycolysis Inhibits Mineralization of Cementoblasts Under Compression by Suppressing the Piezo1/Wnt Signaling

doi: 10.7150/ijms.109287

Figure Lengend Snippet: HK2-mediated glycolysis inhibited cementoblasts' mineralization by suppressing Piezo1 expression. (A) The knockdown efficiency of the three HK2 siRNAs was verified by qRT-PCR and WB. (B) Glucose consumption and lactate production in cementoblasts, normalized to cell number. (C) and (D) The relative mRNA and protein levels of Piezo1, RANKL, OPG, RANKL/OPG, Runx2, OSX, and OPN were identified by qRT-PCR and WB. The blot images represented data from 3 independent experiments. (E) Immunofluorescence of Piezo1 with DAPI counterstaining. Scale bar = 20μm. (F) Von Kossa (VK), Alizarin Red S (ARS) (after 3 weeks of mineralization induction) and alkaline phosphatase staining (after 2 weeks of mineralization induction). Scale bar = 10μm. The data are presented as mean ± SD (*P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001). Con: Control, F: Force.

Article Snippet: The PVDF membranes were blocked at room temperature with 5% bovine serum albumin (BSA) for 1 hour, followed by incubation overnight at 4°C with primary antibodies against Piezo1 (1:1000, ABclonal, China), GLUT1 (1:5000, ABclonal, China), HK2 (1:1000, Abcam, UK), PFKFB3 (1:2000, Abcam, UK), LDHA (1:2000, Abcam, UK), β-catenin (1:1000, ABclonal, China), TCF1 (1:1000, ABclonal, China), LEF1 (1:1000, ABclonal, China), RANKL (1:1000, Proteintech, USA), OPG (1:1000, Proteintech, USA), Runx2 (1:1000, Cell Signaling Technology, USA), OSX (1:1000, Cell Signaling Technology, USA), and OPN (1:1500, Cell Signaling Technology, USA), all from rabbit sources.

Techniques: Expressing, Knockdown, Quantitative RT-PCR, Immunofluorescence, Staining, Control

Journal: Journal of Oral Biology and Craniofacial Research

Article Title: The effect of vitamin D deficiency on the RANKL/OPG ratio in rats

doi: 10.1016/j.jobcr.2022.02.004

Figure Lengend Snippet: Assay Kits used in the study.

Article Snippet: Rat OPG (Osteoprotegerin) ELISA Kit , E-EL-R0050 96T , Serum OPG detection , Elabscience biotechnology Inc, China.

Techniques: Colorimetric Assay, Calcium Assay, Enzyme-linked Immunosorbent Assay