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: Cellular Reprogramming

Article Title: Improvement of Chicken Primordial Germ Cell Maintenance In Vitro by Blockade of the Aryl Hydrocarbon Receptor Endogenous Activity

doi: 10.1089/cell.2016.0015

Figure Lengend Snippet: Alpha-naphthoflavone increases the number of cells with ALP activity in cultured PGCs. PGCs were cultured as described in the Materials and Methods section and maintained in control medium (left panel) or in medium containing α-naphthoflavone (aNF, 12 μM, right panel) for an additional period of 10 days. Alkaline phosphatase staining was performed as described in the Materials and Methods section to identify undifferentiated primordial cells. Photographs shown are representative images of one of the three independent experiments performed. Lower panel: Intensity of staining, expressed in arbitrary units as the mean ± SEM of three independent experiments, of cultures maintained in control medium or in medium containing α-naphthoflavone (aNF). aSignificant difference between the staining intensity of control and treated cultures, p < 0.05. PGCs, primordial germ cells. Color images available online at www.liebertpub.com/cell

Article Snippet: Cultured PGCs were fixed in 4% neutral paraformaldehyde for 20 minutes, and alkaline phosphatase (ALP) staining was performed according to the conventional histochemical method with NBT/BCIP (Promega) as the substrate and following the manufacturer's instructions.

Techniques: Activity Assay, Cell Culture, Control, Staining