Journal: Bioengineering

Article Title: An Injectable, Osteoconductive Gelatin-Enabled GelMA/HAp Hydrogel Scaffold for Minimally Invasive Bone Tissue Engineering

doi: 10.3390/bioengineering13020139

Figure Lengend Snippet: ( a ) Immunofluorescence images of Runx2 in BMSCs co-cultured with hydrogels during osteogenic differentiation. ( b ) Number of Runx2-positive cells in BMSCs co-cultured with hydrogels. *** p < 0.005; **** p < 0.0001.

Article Snippet: The fixed samples were permeabilized and blocked, followed by incubation with a primary antibody against Runx2 (Shanghai, China, Proteintech, 20700-1-AP) and then with the corresponding fluorophore-conjugated secondary antibody (USA, Thermo Fisher Scientific, A-11011).

Techniques: Immunofluorescence, Cell Culture

Journal: Bioengineering

Article Title: An Injectable, Osteoconductive Gelatin-Enabled GelMA/HAp Hydrogel Scaffold for Minimally Invasive Bone Tissue Engineering

doi: 10.3390/bioengineering13020139

Figure Lengend Snippet: ( a – d ) qPCR analysis of OCN , COL-1 , Runx2 and Osterix in BMSCs on Day 7 and 14. * p < 0.05; **** p < 0.0001.

Article Snippet: The fixed samples were permeabilized and blocked, followed by incubation with a primary antibody against Runx2 (Shanghai, China, Proteintech, 20700-1-AP) and then with the corresponding fluorophore-conjugated secondary antibody (USA, Thermo Fisher Scientific, A-11011).

Techniques:

Journal: Journal of tissue engineering

Article Title: Extracellular vesicles derived from human dental pulp stem cells promote osteogenesis of adipose-derived stem cells via the MAPK pathway.

doi: 10.1177/2041731420975569

Figure Lengend Snippet: Figure 1. HDPSCs improved the osteogenic differentiation of hADSCs, probably via EVs. HDPSCs and hADSCs were co-cultured in the presence of GW4869 or DMSO. (a) ALP staining of the co-cultured hDPSCs and hADSCs. (b) Quantitative analysis of ALP activity in the co-cultured hDPSCs and hADSCs. (c) Alizarin Red S staining of the co-cultured hDPSCs and hADSCs. Calcium nodules are indicated by white arrows. (d, e) PCR detection of BMP2 and RUNX2 expression involved in osteogenic differentiation of co-cultured hDPSCs and hADSCs. Data are presented as the mean ± standard deviation of the mean (n = 3), * represents p < 0.05, ** represents p < 0.01.

Article Snippet: Exosome-mediated stem cell differentiation in vitro To evaluate the effect of hDPSC-EVs on osteogenic differentiation of hADSCs, we performed ALP staining, ALP activity assays, qRT-PCR and immunofluorescence assays (IFAs) of RUNX2 and osteocalcin (OCN), as previously described.26,27 The following four groups were assessed: (1) hADSCs cultured in osteoblast-inducing medium (OM) (Cyagen, Santa Clara, CA, USA) alone (0 μg/mL); (2) hADSCs cultured in OM supplemented with 10 μg/mL hDPSC-EVs (10 μg/mL); (3) hADSCs cultured in OM supplemented with 20 μg/mL hDPSC-EVs (20 μg/mL); and (4) hADSCs cultured in OM supplemented with 40 μg/mL hDPSC-EVs (40 μg/mL).

Techniques: Cell Culture, Staining, Activity Assay, Expressing, Standard Deviation

Journal: Journal of tissue engineering

Article Title: Extracellular vesicles derived from human dental pulp stem cells promote osteogenesis of adipose-derived stem cells via the MAPK pathway.

doi: 10.1177/2041731420975569

Figure Lengend Snippet: Figure 3. The effects of hDPSC-EVs on the migration and osteogenic differentiation of hADSCs. (a) Transwell migration assay to detect the effect of hDPSC-EVs on the migration of hADSCs. (b) Quantitative analysis of the effects of hDPSC-EVs on the migration of hADSCs. (c) ALP staining to detect the effect of hDPSC-EVs on osteogenic differentiation of hADSCs. (d) Quantitative detection of ALP activity to detect the effects of hDPSC-EVs on osteogenic differentiation of hADSCs. (e, f) The effects of hDPSC- EVs on the expression of the osteogenic-related genes RUNX2 and BMP2 in hADSCs. (g, h) Immunofluorescence detection of RUNX2 and OCN expression in hADSCs after 7 days incubation with 40 µg/mL hDPSC-EVs. Data are presented as the mean ± standard deviation of the mean (n = 3), * represents p < 0.05, ** represents p < 0.01.

Article Snippet: Exosome-mediated stem cell differentiation in vitro To evaluate the effect of hDPSC-EVs on osteogenic differentiation of hADSCs, we performed ALP staining, ALP activity assays, qRT-PCR and immunofluorescence assays (IFAs) of RUNX2 and osteocalcin (OCN), as previously described.26,27 The following four groups were assessed: (1) hADSCs cultured in osteoblast-inducing medium (OM) (Cyagen, Santa Clara, CA, USA) alone (0 μg/mL); (2) hADSCs cultured in OM supplemented with 10 μg/mL hDPSC-EVs (10 μg/mL); (3) hADSCs cultured in OM supplemented with 20 μg/mL hDPSC-EVs (20 μg/mL); and (4) hADSCs cultured in OM supplemented with 40 μg/mL hDPSC-EVs (40 μg/mL).

Techniques: Migration, Transwell Migration Assay, Staining, Activity Assay, Expressing, Immunofluorescence, Incubation, Standard Deviation

Journal: Cell reports

Article Title: Mapping Distinct Bone Marrow Niche Populations and Their Differentiation Paths

doi: 10.1016/j.celrep.2019.06.031

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: shRNA-TF510502 (Runx2) , Origene , 4 unique shRNA.

Techniques: Recombinant, shRNA, Construct, Plasmid Preparation, Sequencing, Software

Journal: bioRxiv

Article Title: The unique function of Runx1 in skeletal muscle differentiation and regeneration is mediated by an ETS interaction domain

doi: 10.1101/2023.11.21.568117

Figure Lengend Snippet: (A) Western blot analysis of Runx1and Runx2 in WT and Runx1KO C2C12 relative to β-actin. (B) Representative phase contrast microscopy images of WT and Runx1KO C2C12 cells in growth medium (GM) or after 6 d in differentiation medium (DM). (C) Quantification of fusion index of WT and Runx1KO C2C12 cells after 6 days in differentiation medium. ***p<0.001. Data present as means ± SEM. n=3 for each group. (D) Differential gene expression analysis reveal differences in GM (D0) and DM (D1,2,6). The affected genes in each group/day are displayed here and in Supplemental Table S1. (E) Heat maps of the changes in 368 transcription factors: above the clusters, relative Log2 fold change in each column is shown. Below the clusters, the relative abundance (LogCPM) is shown. Members of the Notch pathway, Ets, MRF gene families are indicated above their relative position. (F) GO terms of the up- and downregulated genes in Runx1KO. Details in Supplemental Table S1.

Article Snippet: The mouse Runx2 plasmid pCMV-Flag-mRunx2 was purchased from Origene.

Techniques: Western Blot, Microscopy, Gene Expression

Journal: bioRxiv

Article Title: The unique function of Runx1 in skeletal muscle differentiation and regeneration is mediated by an ETS interaction domain

doi: 10.1101/2023.11.21.568117

Figure Lengend Snippet: (A) Runx1KO C2C12 cells were transfected with 200ng Runx1 or Runx2 plasmids. After 24 hr, cells were induced to differentiate for 6 days followed by immunofluorescence staining for MHC (red) and DNA (Hoechst, blue). (B) Quantification of fusion index and of (C) MHC+ cell lengths in experiments shown in A. (D) Schematics of Runx1 and Runx2 chimera proteins. (E) Quantification of fusion index (F) and MHC+ cell lengths in experiments shown in G; (G) Runx1KO C2C12 cells were transfected with 200ng of five different chimera plasmids numbered in D. 24 h for after transfection, cells were induced to differentiate for 6 days followed by immunofluorescence staining for MHC (red) and DNA (Hoechst, Blue). Data presented as means ± SEM. n=3 for each group. ***p<0.001. n.s., not significant (Scale bars: 150μm.).

Article Snippet: The mouse Runx2 plasmid pCMV-Flag-mRunx2 was purchased from Origene.

Techniques: Transfection, Immunofluorescence, Staining

Journal: bioRxiv

Article Title: The unique function of Runx1 in skeletal muscle differentiation and regeneration is mediated by an ETS interaction domain

doi: 10.1101/2023.11.21.568117

Figure Lengend Snippet: (A) Western blots from co-immunoprecipitation experiments in C2C12 cells transfected with Myc tagged Runx proteins and Flag-tagged Etv4. (B) Venn diagram of ATACseq peaks from C2C12 or Runx1KO cells. Most peaks are shared, 9,318 peaks that are only found to C2C12 and 2,607 peaks are only found in Runx1KO C2C12 cells. (C) Heatmaps showing ATACseq reads from WT or Runx1KO C2C12 cells mapped onto these 3 classes of ATACseq peaks (WT specific, Shared, and Runx1KO specific). (D) Graph displaying the -log pValue of transcription factor motif enrichment as determined by HOMER. Runx1 motifs (blue) are more highly enriched in WT C2C12 cells, whereas Mef2 motifs (red MADS-box) are more enriched in Runx1KO C2C12. The inset shows that AP1 motifs (purple) are similarly enriched in both WT and C2C12 cells as expected for a global enhancer binding factor. (E) Bar graph showing enrichment for the Ets:Runx composite motif in Runx1 ChIP data from myoblasts or T-ALL cells, less enrichment seen in Runx2 ChIP from preosteoblasts. The composite motif enrichment in each dataset is normalized to enrichment rate of the Runx-only motif in the respective ChIP from each cell types.

Article Snippet: The mouse Runx2 plasmid pCMV-Flag-mRunx2 was purchased from Origene.

Techniques: Western Blot, Immunoprecipitation, Transfection, Binding Assay

Journal: Journal of the mechanical behavior of biomedical materials

Article Title: Biodegradable and osteoconductive sodium alginate-gelatin/amorphous magnesium phosphate 3D-printed scaffolds for craniofacial bone regeneration

doi: 10.1016/j.jmbbm.2025.107284

Figure Lengend Snippet: COL1A1 , ALPL , RUNX2 , and OCN expression of alveolar bone-derived mesenchymal stem cells (aBMSCs) cultured on 3D-printed hydrogel scaffolds, analyzed by RT-qPCR at 7, 14, and 21 days. Data are presented as mean ± SD. Different uppercase letters denote significant differences between the groups (One-way ANOVA/Sidak, n = 6, α = 5 %).

Article Snippet: Next, 500 ng RNA was reversed to cDNA by SuperScript II Reverse Transcriptase (Invitrogen), followed by qPCR reactions using standard reagents (TaqMan Gene Expression Master Mix; Thermo Fisher Scientific) to amplify the gene expression levels of collagen type 1 ( COL1A1 , Hs01076756_g1), alkaline phosphatase ( ALPL , Hs01029144_m1) osteocalcin ( OCN ; Hs01587814_g1), and Runt-related transcription factor 2 ( RUNX2 ; Hs01047973_m1).

Techniques: Expressing, Derivative Assay, Cell Culture, Quantitative RT-PCR