Journal: Journal of Orthopaedic Surgery and Research

Article Title: KLF5 promotes the ossification process of ligamentum flavum by transcriptionally activating CX43

doi: 10.1186/s13018-024-04702-2

Figure Lengend Snippet: KLF5 expression was upregulated in OLF fibroblasts. (A) A flow diagram of primary isolation of clinical samples. (B) KLF5 expression in OLF fibroblasts and normal LF fibroblasts was detect by qRT-PCR analysis. (C) IF staining was used to detect the KLF5 expression in Normal-LF fibroblasts and OLF fibroblasts (scale bar = 100 μm). (D) Western blot was used to detect the protein of KLF5 expression. (E) ALP activity was measured using ALP staining (scale bar = 100 μm); (F) osteogenic differentiation of OLF fibroblasts was analyzed using ARS staining (scale bar = 100 μm). (G) Levels of osteogenic specific markers (RUNX2, OPN, and OCN) were detected using western blot. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The detection of ALP activity was performed using an ALP staining kit (Stemgent 00–0055; Mito Biological technology Co,.LTD, Shanghai, China) following the manufacturer’s instructions, then washed with PBS and examined under a microscope to visualize ALP activity.

Techniques: Expressing, Isolation, Quantitative RT-PCR, Staining, Western Blot, Activity Assay

Journal: Journal of Orthopaedic Surgery and Research

Article Title: KLF5 promotes the ossification process of ligamentum flavum by transcriptionally activating CX43

doi: 10.1186/s13018-024-04702-2

Figure Lengend Snippet: KLF5 knockdown significantly decreased the osteogenic ability of OLF fibroblasts (A) The silencing effect of siRNA on KLF5 was measured using western blot in OLF fibroblasts. (B) ALP activity was measured using ALP staining (scale bar = 100 μm). (C) Osteogenic differentiation of OLF fibroblasts was measured using ARS staining (scale bar = 100 μm). (D) Levels of osteogenic specific markers (RUNX2, OPN, and OCN) were detected using western blot. ** p < 0.01, *** p < 0.001

Article Snippet: The detection of ALP activity was performed using an ALP staining kit (Stemgent 00–0055; Mito Biological technology Co,.LTD, Shanghai, China) following the manufacturer’s instructions, then washed with PBS and examined under a microscope to visualize ALP activity.

Techniques: Knockdown, Western Blot, Activity Assay, Staining

Journal: Journal of Orthopaedic Surgery and Research

Article Title: KLF5 promotes the ossification process of ligamentum flavum by transcriptionally activating CX43

doi: 10.1186/s13018-024-04702-2

Figure Lengend Snippet: Overexpression of KLF5 significantly increased the osteogenic ability of normal LF fibroblasts. (A) The effect of lenti-KLF5 in normal LF fibroblasts was measured using western blot. (B) ALP activity was measured using ALP staining (scale bar = 100 μm). (C) Osteogenic differentiation of normal-LF fibroblasts was measured using ARS staining (scale bar = 100 μm). (D) Levels of osteogenic specific markers (RUNX2, OPN, and OCN) were detected using western blot. ** p < 0.01, *** p < 0.001

Article Snippet: The detection of ALP activity was performed using an ALP staining kit (Stemgent 00–0055; Mito Biological technology Co,.LTD, Shanghai, China) following the manufacturer’s instructions, then washed with PBS and examined under a microscope to visualize ALP activity.

Techniques: Over Expression, Western Blot, Activity Assay, Staining

Journal: Journal of Orthopaedic Surgery and Research

Article Title: KLF5 promotes the ossification process of ligamentum flavum by transcriptionally activating CX43

doi: 10.1186/s13018-024-04702-2

Figure Lengend Snippet: KLF5 targeted CX43 to promote osteogenic differentiation of OLF fibroblasts. (A) Protein expression levels of CX43 after OLF fibroblasts after the intervention of KLF5 and overexpression of CX43. (B) ALP activity was measured using ALP staining (scale bar = 100 μm). (C) Osteogenic differentiation of OLF fibroblasts was measured using ARS staining (scale bar = 100 μm). (D) Levels of osteogenic specific markers (RUNX2, OPN, and OCN) were detected using western blot. * p < 0.05, *** p < 0.001

Article Snippet: The detection of ALP activity was performed using an ALP staining kit (Stemgent 00–0055; Mito Biological technology Co,.LTD, Shanghai, China) following the manufacturer’s instructions, then washed with PBS and examined under a microscope to visualize ALP activity.

Techniques: Expressing, Over Expression, Activity Assay, Staining, Western Blot

Journal: Cells

Article Title: Topical Application of Butyl Flufenamate Ointment Promotes Cranial Defect Healing in Mice by Inducing BMP2 Secretion in Skin Mesenchymal Stem Cells

doi: 10.3390/cells11223620

Figure Lengend Snippet: Isolation and stemness determination of mSMSCs. ( a ) Images of mouse mesenchymal stem cells. Scale bar = 500 μm. ( b ) ALP staining and ARS staining after osteogenic induction of mSMSCs. ( c ) Oil red O staining after adipogenic induction of mSMSCs. Scale bar = 200 μm. ( d ) Alcian blue staining after chondrogenic induction.

Article Snippet: An ALP staining kit (CWBIO, Beijing, China) was used to assess ALP expression, and the images of stained cells were then scanned.

Techniques: Isolation, Staining

Journal: Cells

Article Title: Topical Application of Butyl Flufenamate Ointment Promotes Cranial Defect Healing in Mice by Inducing BMP2 Secretion in Skin Mesenchymal Stem Cells

doi: 10.3390/cells11223620

Figure Lengend Snippet: Co-culture with mSMSCs promoted osteogenesis of mBMMSCs in the presence of a low concentration of FFA. ( a ) The process of obtaining mSMSCs and the pattern of mSMSC and mBMMSC growth in co-culture. ( b ) Cell proliferation values for two groups, obtained using the Cell Counting Kit-8 on days 1, 2, 3, 4, 5, 6, and 7. ( c ) ALP staining and quantification showing that co-culture with mSMSCs markedly increased ALP activity in mBMMSCs in the presence of FFA. ( d ) ARS staining and quantification showing that co-culture with mSMSCs significantly accelerated mBMMSC mineralization in the presence of FFA. ( e ) qRT-PCR showing that co-culture with mSMSCs increased the mRNA expression of runx2 , bglap , and osx in mBMMSCs in the presence of FFA. ** p < 0.01, *** p < 0.001.

Article Snippet: An ALP staining kit (CWBIO, Beijing, China) was used to assess ALP expression, and the images of stained cells were then scanned.

Techniques: Co-Culture Assay, Concentration Assay, Cell Counting, Staining, Activity Assay, Quantitative RT-PCR, Expressing