antibody against grem2  (Boster Bio)

Journal: Hereditas

Article Title: Overexpression of miR-671-3p alleviates postmenopausal osteoporosis by targeting GREM2 to activate BMP2/SMAD signaling pathway.

doi: 10.1186/s41065-025-00467-8

Figure Lengend Snippet: Fig. 3 WT/MT-GREM2 3’UTR sequences and miR-671-3p binding sites (A). Dual-luciferase reporter gene assay (B) and RNA immunoprecipitation (RIP) experiment (C) indicated the binding between miR-671-3p and GREM2. The expression of GREM2 was upregulated in PMOP patients’ serum (D) and negatively related to the levels of miR-671-3p (E). GREM2 was upregulated in MG, and overexpression of miR-671-3p inhibited its expression (F). GREM2 overexpression had no impact on miR-671-3p expression (G). WT-GREM2: Wild-type GREM2 3’UTR luciferase reporter; MT-GREM2: Mutant GREM2 3’UTR luciferase reporter; Ctrl: Normal culture conditions; MG: Cells treated with simulated MG for 72 h; MG + miR-NC: MG + miR- NC transfection; MG + miR- mimic: MG + miR-671-3p mimic transfection; MG + miR + GREM2-NC: MG + miR-mimic + empty vector transfection; MG + miR + oe-GREM2: MG + miR- mimic + GREM2 overexpression plasmid transfection. Bar graphs show mean ± SD (n = 5). two-tailed t-test for WT vs. MT; one-way ANOVA with Tukey’s test for other groups

Article Snippet: Proteins were detected using antibodies against GREM2 (1:1000, Proteintech), BMP2 (1:1000, CST), p-SMAD1/5 (1:1000, Abcam), and GAPDH (1:5000, Proteintech).

Techniques: Binding Assay, Luciferase, Reporter Gene Assay, RNA Immunoprecipitation, Expressing, Over Expression, Mutagenesis, Transfection, Plasmid Preparation, Two Tailed Test

Journal: Hereditas

Article Title: Overexpression of miR-671-3p alleviates postmenopausal osteoporosis by targeting GREM2 to activate BMP2/SMAD signaling pathway.

doi: 10.1186/s41065-025-00467-8

Figure Lengend Snippet: Fig. 4 miR-671-3p inhibits GREM2 protein expression (A). Elevated GREM2 exacerbates MG-induced inhibition of cell differentiation (B-C), disrupted vi ability (D) and apoptosis (E-F), counteracting the protective effect of miR-671-3p. Ctrl: Normal culture conditions; MG: Cells treated with simulated MG for 72 h; MG + miR-NC: MG + miR- NC transfection; MG + miR-mimic: MG + miR-671-3p mimic transfection; MG + miR + GREM2-NC: MG + miR-mimic + empty vector transfection; MG + miR + oe-GREM2: MG + miR-mimic + GREM2 overexpression plasmid transfection. Bar graphs show mean ± SD (n = 5). one-way ANOVA with Tukey’s post hoc test

Article Snippet: Proteins were detected using antibodies against GREM2 (1:1000, Proteintech), BMP2 (1:1000, CST), p-SMAD1/5 (1:1000, Abcam), and GAPDH (1:5000, Proteintech).

Techniques: Expressing, Inhibition, Cell Differentiation, Transfection, Plasmid Preparation, Over Expression

Journal: Hereditas

Article Title: Overexpression of miR-671-3p alleviates postmenopausal osteoporosis by targeting GREM2 to activate BMP2/SMAD signaling pathway.

doi: 10.1186/s41065-025-00467-8

Figure Lengend Snippet: Fig. 5 Reduced expression of BMP2 (A) and P-SMAD1/5 (B-C) proteins under MG is promoted by miR-671-3p upregulation and inhibited by GREM2 overexpression. Ctrl: Normal culture conditions; MG: Cells treated with simulated MG for 72 h; MG + miR-NC: MG + miR- NC transfection; MG + miR-mimic: MG + miR-671-3p mimic transfection; MG + miR + GREM2-NC: MG + miR-mimic + empty vector transfection; MG + miR + oe-GREM2: MG + miR-mim ic + GREM2 overexpression plasmid transfection. Bar graphs show mean ± SD (n = 5). one-way ANOVA with Tukey’s post hoc test

Article Snippet: Proteins were detected using antibodies against GREM2 (1:1000, Proteintech), BMP2 (1:1000, CST), p-SMAD1/5 (1:1000, Abcam), and GAPDH (1:5000, Proteintech).

Techniques: Expressing, Over Expression, Transfection, Plasmid Preparation

Journal: Cellular & Molecular Biology Letters

Article Title: LncRNA DNM3OS regulates GREM2 via miR-127-5p to suppress early chondrogenic differentiation of rat mesenchymal stem cells under hypoxic conditions

doi: 10.1186/s11658-021-00269-6

Figure Lengend Snippet: Primer sequences used for RT-qPCR

Article Snippet: After finishing blocking in 0.1% BSA, the rMSCs were incubated with the primary antibody against GREM2 overnight at 4 °C, followed by incubation with CY3-conjugated goat anti-rabbit IgG (1:100 dilution; Boster Biological Technology) for 1 h at room temperature.

Techniques:

Journal: Cellular & Molecular Biology Letters

Article Title: LncRNA DNM3OS regulates GREM2 via miR-127-5p to suppress early chondrogenic differentiation of rat mesenchymal stem cells under hypoxic conditions

doi: 10.1186/s11658-021-00269-6

Figure Lengend Snippet: GREM2 was a target of miR-127-5p. A Nucleotide sequences of the predicted target site for miR-127-5p in the GREM2 3ʹ-UTR. B Luciferase reporter assays were performed to identify direct interaction between miR-127-5p and the GREM2 3ʹ-UTR. Wild type and mutant miR-127-5p target binding sequences in the GREM2 3ʹ-UTR were cloned into a reporter luciferase vector and co-transfected with the synthetized miR-127-5p (or NC) into rMSCs. *** p < 0.001, compared with NC; C GREM2 protein expression was measured in rMSCs from the control, induction, hypoxia, and hypoxia/induction groups. D GREM2 protein expression was measured in rMSCs from the control, induction, and si-SHP/hypoxia groups. E GREM2 protein expression was measured in rMSCs from the control, induction, and miR-127-5p inhibitor/induction groups. F GREM2 protein expression in rMSCs was measured by immunofluorescence. * p < 0.05, *** p < 0.001, compared with control; # p < 0.05, ### p < 0.001, compared with hypoxia or induction group

Article Snippet: After finishing blocking in 0.1% BSA, the rMSCs were incubated with the primary antibody against GREM2 overnight at 4 °C, followed by incubation with CY3-conjugated goat anti-rabbit IgG (1:100 dilution; Boster Biological Technology) for 1 h at room temperature.

Techniques: Luciferase, Mutagenesis, Binding Assay, Clone Assay, Plasmid Preparation, Transfection, Expressing, Control, Immunofluorescence

Journal: Cellular & Molecular Biology Letters

Article Title: LncRNA DNM3OS regulates GREM2 via miR-127-5p to suppress early chondrogenic differentiation of rat mesenchymal stem cells under hypoxic conditions

doi: 10.1186/s11658-021-00269-6

Figure Lengend Snippet: Overexpression of miR-127-5p promoted chondrogenic differentiation of rMSCs by regulating GREM2-mediated SMAD-dependent BMP signaling. The rMSCs were transfected with miR-127-5p mimics, followed by hypoxia intervention (1% O 2 ) and induction of chondrogenic differentiation for 14 days. A MiR-127-5p and GREM2 expression levels were determined by quantitative real-time PCR analysis. Results are expressed as mean ± standard deviation of data obtained from three independent experiments. *** p < 0.001, compared with control; ### p < 0.001, compared with induction group; p < 0.001, compared with hypoxia/induction; B Images from Alcian blue staining assays of rMSCs. C Levels of COL1A1, COL2A1, SOX9, and ACAN protein expression were detected in rMSCs. D Levels of GREM2, BMP2, p-SMAD1, and p-SMAD2 protein expression in rMSCs were detected by western blotting. E GREM2 protein expression in rMSCs was detected by immunofluorescence

Article Snippet: After finishing blocking in 0.1% BSA, the rMSCs were incubated with the primary antibody against GREM2 overnight at 4 °C, followed by incubation with CY3-conjugated goat anti-rabbit IgG (1:100 dilution; Boster Biological Technology) for 1 h at room temperature.

Techniques: Over Expression, Transfection, Expressing, Real-time Polymerase Chain Reaction, Standard Deviation, Control, Staining, Western Blot, Immunofluorescence

Journal: Cellular & Molecular Biology Letters

Article Title: LncRNA DNM3OS regulates GREM2 via miR-127-5p to suppress early chondrogenic differentiation of rat mesenchymal stem cells under hypoxic conditions

doi: 10.1186/s11658-021-00269-6

Figure Lengend Snippet: DNM3OS inhibited chondrogenic differentiation of rMSCs via miR-127-5p regulation of GREM2. The rMSCs were transfected with a DNM3OS overexpression plasmid, followed by induction of chondrogenic differentiation for 14 days. Accordingly, the rMSCs were classified into control, induction, hypoxia, hypoxia/induction, and DNM3OS/induction groups. A The putative binding site for miR-127-5p in the DNM3OS 3′-UTR is shown. B DNM3OS expression was determined by quantitative real-time PCR analysis. C A luciferase reporter assay was performed to identify direct action between DNM3OS and the GREM2 3ʹ-UTR. D Levels of DNM3OS, miR-127-5p, and GREM2 expression were determined by quantitative real-time PCR analysis. Results are expressed as mean ± standard deviation of data obtained from three independent experiments. ** p < 0.01, *** p < 0.001, compared with control; # p < 0.05, ### p < 0.001, compared with induction group; E Images from Alcian blue staining assays of rMSCs. (F) Levels of COL1A1, COL2A1, SOX9, and ACAN protein expression were detected in rMSCs. G Levels of GREM2, BMP2, p-SMAD1, and p-SMAD2 in rMSCs were detected by western blotting. H GREM2 protein expression in rMSCs was measured by immunofluorescence

Article Snippet: After finishing blocking in 0.1% BSA, the rMSCs were incubated with the primary antibody against GREM2 overnight at 4 °C, followed by incubation with CY3-conjugated goat anti-rabbit IgG (1:100 dilution; Boster Biological Technology) for 1 h at room temperature.

Techniques: Transfection, Over Expression, Plasmid Preparation, Control, Binding Assay, Expressing, Real-time Polymerase Chain Reaction, Luciferase, Reporter Assay, Standard Deviation, Staining, Western Blot, Immunofluorescence

Journal: Cellular & Molecular Biology Letters

Article Title: LncRNA DNM3OS regulates GREM2 via miR-127-5p to suppress early chondrogenic differentiation of rat mesenchymal stem cells under hypoxic conditions

doi: 10.1186/s11658-021-00269-6

Figure Lengend Snippet: DNM3OS knockdown weakened the inhibitory effect of hypoxia on chondrogenic induction. The rMSCs were transfected with si-DNM3OS, followed by induction of chondrogenic differentiation under hypoxic conditions (1% O 2 ) for 14 days. A Levels of miR-127-5p, DNM3OS, and GREM2 expression were determined by quantitative real-time PCR analysis. *** p < 0.001, compared with si-NC + hypoxia/induction group. B Levels of GREM2, BMP2, p-SMAD1, and p-SMAD2 in rMSCs were detected by western blotting. C Images from Alcian blue staining assays of rMSCs. (D) Levels of COL1A1, COL2A1, SOX9, and ACAN protein expression were detected in rMSCs. si-NC: negative control

Article Snippet: After finishing blocking in 0.1% BSA, the rMSCs were incubated with the primary antibody against GREM2 overnight at 4 °C, followed by incubation with CY3-conjugated goat anti-rabbit IgG (1:100 dilution; Boster Biological Technology) for 1 h at room temperature.

Techniques: Knockdown, Transfection, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Staining, Negative Control