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: International Journal of Molecular Sciences

Article Title: Myogenetic Oligodeoxynucleotide Induces Myocardial Differentiation of Murine Pluripotent Stem Cells

doi: 10.3390/ijms241814380

Figure Lengend Snippet: iSN04 inhibits proliferation of undifferentiated miPSCs and mESCs. ( A ) Representative fluorescence images of 20D17 cells treated with 10 μM iSN04 in GM for 24 and 48 h. Scale bar, 200 μm. Nanog -GFP + colony size was quantified. * p < 0.05 vs. control (Student’s t -test). n = 4 fields. ( B ) qPCR results of 20D17 cells treated with 10 μM iSN04 in GM for 48 h. n = 3. ( C ) Representative images of ALP staining of hCGp7 cells treated with 10 μM iSN04 in GM for 48 h. Scale bar, 200 μm. ALP + colony size was quantified. ** p < 0.01 vs. control (Student’s t -test). n = 4 fields.

Article Snippet: ALP enzymatic activity of the hCGp7 cell colonies was visualized using an ALP Stain Kit (Fujifilm Wako Chemicals) according to the manufacturer’s instructions [ ].

Techniques: Fluorescence, Staining

Journal: Molecular Medicine Reports

Article Title: The COL6A1 rs201153092 single nucleotide polymorphism, associates with thoracic ossification of the posterior longitudinal ligament

doi: 10.3892/mmr.2019.10846

Figure Lengend Snippet: COL6A1 gene rs201153092A site mutation induces osteogenic differentiation of 3T3-E1 cells. After 21 days of osteogenic induction, (A) ALP staining and (B) Alizarin red staining were performed in the COL6A1 gene mutation, non-mutation, control and mock groups. ***P<0.001 vs. COL6A1-rs201153092A. mRNA expression levels of (C) COL6A1 and (D) ALP in the COL6A1 gene mutation and non-mutation groups were normalized to the control group. ***P<0.001 vs. COL6A1-rs201153092A. ALP, alkaline phosphatase; COL6A1, collagen 6A1; Ctrl, control.

Article Snippet: Osteogenic identification was assessed using an ALP activity staining kit (cat. no. GMS80033.1; Genmed Scientifics, Inc.) as in our previous study ( ).

Techniques: Mutagenesis, Staining, Control, Expressing