mouse pre osteoblast cell line  (ATCC)

Journal: Genes & Diseases

Article Title: Targeting MDK alleviates bone loss via dual regulation of osteogenic differentiation and inflammatory cytokine expression

doi: 10.1016/j.gendis.2025.101931

Figure Lengend Snippet: Recombinant MDK protein inhibits osteogenic differentiation in vitro in a dose-dependent manner. (A) Cell viability after treating MC3T3-E1 cells with recombinant MDK protein after 48 h, assessed using the CCK-8 assay. Inter-group comparisons were analyzed by one-way ANOVA. (B) Western blotting analysis of ALP, RUNX2, OSX, and OCN expression levels following MDK treatment (7 days). (C–F) Reverse transcription PCR analysis of mRNA expression levels of Alpl , Runx2 , Sp7 , and Bglap in MC3T3-E1 cells following MDK treatment (7 days). β-actin served as the internal control. Inter-group comparisons were analyzed by one-way ANOVA. (G, H) ALP staining and activity assays were performed after inducing MC3T3-E1 cells with recombinant MDK protein (0–600 ng/mL) for 14 days. Inter-group comparisons were analyzed by one-way ANOVA. (I, J) ARS staining and quantitative analysis were conducted after inducing MC3T3-E1 cells with recombinant MDK protein (0–600 ng/mL) for 21 days. Inter-group comparisons were analyzed by one-way ANOVA. Scale bar, 100 μm ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001; “ns” indicates non-significant differences.

Article Snippet: Procell Life Science & Technology Co., Ltd. (Wuhan, China) supplied the pre-osteoblastic cell line MC3T3-E1 Subclone 14.

Techniques: Recombinant, In Vitro, CCK-8 Assay, Western Blot, Expressing, Reverse Transcription, Control, Staining, Activity Assay

Journal: Genes & Diseases

Article Title: Targeting MDK alleviates bone loss via dual regulation of osteogenic differentiation and inflammatory cytokine expression

doi: 10.1016/j.gendis.2025.101931

Figure Lengend Snippet: MDK suppresses osteoblast differentiation via the PI3K/AKT signaling pathway. (A, B) Western blot detection of the effect of recombinant MDK protein on the protein expression of molecules in the PI3K/AKT signaling pathway during the differentiation of MC3T3-E1 to osteoblasts (7 days). Inter-group comparisons were analyzed by a two-tailed unpaired Student's t -test (for normally distributed data with equal variance). (C, D) ALP, RUNX2, and OSX expression levels were detected by Western blotting. MC3T3-E1 cells were pretreated with 30 μM LY294002. Osteogenic differentiation was induced for 7 days. Inter-group comparisons were analyzed by one-way ANOVA. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001; “ns” indicates non-significant differences.

Article Snippet: Procell Life Science & Technology Co., Ltd. (Wuhan, China) supplied the pre-osteoblastic cell line MC3T3-E1 Subclone 14.

Techniques: Western Blot, Recombinant, Expressing, Two Tailed Test

Journal: Genes & Diseases

Article Title: Targeting MDK alleviates bone loss via dual regulation of osteogenic differentiation and inflammatory cytokine expression

doi: 10.1016/j.gendis.2025.101931

Figure Lengend Snippet: Recombinant MDK protein triggers the activation of inflammatory cytokines through the NF-κB signaling pathway. (A, B) IL-6, TNFα, and IL-1β expression levels were detected using Western blotting. MC3T3-E1 cells were treated with recombinant MDK protein (600 ng/mL). Osteogenic differentiation was induced for 7 days. Inter-group comparisons were analyzed by a two-tailed unpaired Student's t -test (for normally distributed data with equal variance). (C, D) Western blotting analysis of NF-κB signaling pathway molecules in MC3T3-E1 cells treated with recombinant MDK protein for 7 days during osteoblastic differentiation. Inter-group comparisons were analyzed by two-tailed unpaired Student's t -test (for normally distributed data with equal variance). (E, F) IL-6 and IL-1β expression levels were detected using Western blotting. MC3T3-E1 cells were pretreated with 10 μM BAY 11–7082. Osteogenic differentiation was induced for 7 days. Inter-group comparisons were analyzed by one-way ANOVA. ∗ p < 0.05 and ∗∗ p < 0.01.

Article Snippet: Procell Life Science & Technology Co., Ltd. (Wuhan, China) supplied the pre-osteoblastic cell line MC3T3-E1 Subclone 14.

Techniques: Recombinant, Activation Assay, Expressing, Western Blot, Two Tailed Test

Journal: Scientific Reports

Article Title: Understanding of the different roles of Noggin in the Noggin-BMP-2 and Noggin-BMP-9 dimer complexes at the molecular level

doi: 10.1038/s41598-025-33735-8

Figure Lengend Snippet: Initial and final structures of Noggin-BMP-2 and Noggin-BMP-9 after 100 ns MD simulation. Identification of the binding effect of BMP-2 and BMP-9 to BMPR2 in MC3T3-E1 Cells by Noggin using Immunoprecipitation. ( A ) Initial structures of Noggin-BMP-2 and ( B ) Initial structures of Noggin-BMP-9 ( C ) Aligned structures of initial (White color) and final structures of Noggin-BMP-2 (Coral/Blue) ( D )Aligned structures of initial (White) and final structures of Noggin-BMP-9 (Green/Ultraviolet) ( E ) RMSD of Noggin-BMP-2 and Noggin-BMP-9 complexes during the all-atom MD simulation (100 ns). ( F ) RMSF (Root Mean Squared Fluctuation) of Noggin-BMP-2 and Noggin-BMP-9 complexes during the all-atom MD simulation (100 ns). RMSD and RMSF values ​​represent the mean and standard deviation of three independent simulations. ( G ) Western blot analysis using total cell lysates. ( H ) Immunoprecipitation (IP) of BMPR2 followed by immunoblotting (IB) to detect BMP-2 and BMP-9 binding.

Article Snippet: The mouse pre-osteoblastic cell line MC3T3-E1 was purchased from the American Type Culture Collection (ATCC, CRL-2593, Manassas, VA, USA).

Techniques: Binding Assay, Immunoprecipitation, Standard Deviation, Western Blot

Journal: Scientific Reports

Article Title: Understanding of the different roles of Noggin in the Noggin-BMP-2 and Noggin-BMP-9 dimer complexes at the molecular level

doi: 10.1038/s41598-025-33735-8

Figure Lengend Snippet: Identification of binding effect of BMP-2 and BMP-9 to BMPR2 in MC3T3-E1 cells by Noggin. ( A ) BMP-2–treated group, ( B ) BMP-2 + Noggin–treated group, ( C ) BMP-9–treated group, ( D ) BMP-9 + Noggin–treated group. DAPI (blue), anti-BMPR2 (red), and anti-BMP-2 or anti-BMP-9 (green). Merged images show co-localization (yellow). Scale bar: 10 μm; magnification: ×600. ( E ) Western blot of SMAD1/5/9 phosphorylation under the indicated treatments. Top, p-SMAD1/5/9; middle, total SMAD1/5/9; bottom, GAPDH. ( F) Alkaline phosphatase (ALP) activity measured 60 min after BMP addition using a p-nitrophenyl phosphate colorimetric assay (405 nm). Bars indicate mean ± SD ( n ≥ 3). p < 0.0001 vs. NT for all treated groups; asterisks denote significance relative to NT.

Article Snippet: The mouse pre-osteoblastic cell line MC3T3-E1 was purchased from the American Type Culture Collection (ATCC, CRL-2593, Manassas, VA, USA).

Techniques: Binding Assay, Western Blot, Phospho-proteomics, Activity Assay, Colorimetric Assay