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: Frontiers in Endocrinology

Article Title: Calcitonin Induces Bone Formation by Increasing Expression of Wnt10b in Osteoclasts in Ovariectomy-Induced Osteoporotic Rats

doi: 10.3389/fendo.2020.00613

Figure Lengend Snippet: Calcitonin indirectly increases osteoblast mineralization. Osteoblasts were isolated from the calvariae of 1-day-old newborn SD rats and cultured in groups a-f medium as indicated in Materials and Methods. Analysis of osteoblast mineralization was performed by ALP (A) and alizarin red staining (B) . (C) Quantitative results of the experiment shown in (A,B) , and Wnt10b concentration in mediums of each group before cultured with osteoblasts. *Indicates a significant difference ( p < 0.05). Bar = 500 μm.

Article Snippet: Osteoblasts were stained with an ALP staining kit (AK20-COS, COSMO BIO, Tokyo, Japan).

Techniques: Isolation, Cell Culture, Staining, Concentration Assay