mechanical stimulation related protein rhoa  (Proteintech)

Journal: Materials Today Bio

Article Title: Crosslinking stabilization strategy: A novel approach to cartilage-like repair of annulus fibrosus (AF) defects

doi: 10.1016/j.mtbio.2025.101625

Figure Lengend Snippet: The primer sequences used for the qPCR analysis.

Article Snippet: Then, immunohistochemical (IHC) analyses of collagen I (COL1A1) (1:200, bsm-33400M, Bioss, China), collagen II (COL2A1) (1:200, bsm-33409M, Bioss, China), aggrecan (1:200, sc-33695, Santa Cruz, USA), mechanical stimulation signal pathway-related protein RhoA (1:100, sc-418, Santa Cruz, USA), ROCK (1:200, sc-19974, Santa Cruz, USA), and nerve ingrowth protein GAP 43 (1:50, sc-17790, Santa Cruz, USA) were performed.

Techniques:

Journal: Materials Today Bio

Article Title: Crosslinking stabilization strategy: A novel approach to cartilage-like repair of annulus fibrosus (AF) defects

doi: 10.1016/j.mtbio.2025.101625

Figure Lengend Snippet: Immunohistochemical staining and qPCR analysis of rabbit intervertebral disc at 12 w post-modeling. (A) Immunohistochemical staining of COL 1, COL2, and Aggrecan (The red arrow indicates the positive area). (B) Immunohistochemical staining of RhoA, ROCK, SOX 9 and GAP 43. (C) The qPCR analyses for the relative expression of genes (COL 1, Aggrecan, SOX 9 and RhoA). The error bars indicate SD. n = 4 (∗P < 0.05 and ∗∗P < 0.01).

Article Snippet: Then, immunohistochemical (IHC) analyses of collagen I (COL1A1) (1:200, bsm-33400M, Bioss, China), collagen II (COL2A1) (1:200, bsm-33409M, Bioss, China), aggrecan (1:200, sc-33695, Santa Cruz, USA), mechanical stimulation signal pathway-related protein RhoA (1:100, sc-418, Santa Cruz, USA), ROCK (1:200, sc-19974, Santa Cruz, USA), and nerve ingrowth protein GAP 43 (1:50, sc-17790, Santa Cruz, USA) were performed.

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Mechanical stimulation promotes MSCs healing the lesion of intervertebral disc annulus fibrosus

doi: 10.3389/fbioe.2023.1137199

Figure Lengend Snippet: Primers sequences used for qPCR analyses.

Article Snippet: Diaminobenzidine (DAB, ZLI-9018, ZSGB-BIO, China) staining of the main components of annulus fibrosus collagen 1 (COL1) (1:200, ab34710, Abcam, United States), collagen 2 (COL2) (1:200, ab34712, Abcam, United States), mechanical stimulation related protein RhoA (1:100, 10749-1-AP, Proteintech, China) and ROCK1 (1:200, ab97592, Abcam, United States) were conducted according to the product instructions and viewed under a microscope (IX73, Olympus, Japan).

Techniques:

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Mechanical stimulation promotes MSCs healing the lesion of intervertebral disc annulus fibrosus

doi: 10.3389/fbioe.2023.1137199

Figure Lengend Snippet: DAB staining of RhoA and ROCK1 for specimens of different interventions at 5 weeks. RhoA: (A , B) Disc repaired by Fib MSCs and suture has no positive staining in annulus fissure. (C , D) Disc repaired by Fib-T-G MSCs has a continuous structure with positive cell staining in annulus fissure pointed by arrows. ROCK1: (E , F) Disc repaired by Fib MSCs and suture has no positive staining in annulus fissure. (G , H) Disc repaired by Fib-T-G MSCs has a continuous structure with positive cell staining in annulus fissure pointed by arrows.

Article Snippet: Diaminobenzidine (DAB, ZLI-9018, ZSGB-BIO, China) staining of the main components of annulus fibrosus collagen 1 (COL1) (1:200, ab34710, Abcam, United States), collagen 2 (COL2) (1:200, ab34712, Abcam, United States), mechanical stimulation related protein RhoA (1:100, 10749-1-AP, Proteintech, China) and ROCK1 (1:200, ab97592, Abcam, United States) were conducted according to the product instructions and viewed under a microscope (IX73, Olympus, Japan).

Techniques: Staining

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Mechanical stimulation promotes MSCs healing the lesion of intervertebral disc annulus fibrosus

doi: 10.3389/fbioe.2023.1137199

Figure Lengend Snippet: (A) Relative gene (Collagen1, Collagen2, Aggrecan, Mohawk, RhoA, ROCK1, SOX9) expression in different mechanical conditions after 7 days of culture. (B) Matrix of Collagen and Aggrecan quantification in different mechanical conditions after 7 days of culture. The expression level and quantification are expressed as mean ± SD, (n = 3). (C – H) DAB staining of Collagen 1, 2, and Aggrecan for cell culture in different mechanical conditions for 2 weeks. Cell culture differentiation in stable conditions has few positive matrix staining of Collagen 1, 2, and Aggrecan (pointed by arrows), whereas cell culture differentiation in mechanical stimulation conditions has obvious positive matrix staining of Collagen 1, 2, and Aggrecan (pointed by arrows). (I , J) Sirius red staining for cell culture in different mechanical conditions for 2 weeks. Collagen stained by Sirius red is positive but without aligned fibers seen under polarized light (I) ; collagen stained by Sirius red is positive with obviously aligned fibers seen under polarized light (pointed by dotted lines) (J) . * indicates a significant difference between the different culture conditions, p < 0.05.

Article Snippet: Diaminobenzidine (DAB, ZLI-9018, ZSGB-BIO, China) staining of the main components of annulus fibrosus collagen 1 (COL1) (1:200, ab34710, Abcam, United States), collagen 2 (COL2) (1:200, ab34712, Abcam, United States), mechanical stimulation related protein RhoA (1:100, 10749-1-AP, Proteintech, China) and ROCK1 (1:200, ab97592, Abcam, United States) were conducted according to the product instructions and viewed under a microscope (IX73, Olympus, Japan).

Techniques: Expressing, Staining, Cell Culture

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Mechanical stimulation promotes MSCs healing the lesion of intervertebral disc annulus fibrosus

doi: 10.3389/fbioe.2023.1137199

Figure Lengend Snippet: (A) Relative gene expression quantified by qPCR of normal and RhoA gene activated MSCs culture both in differentiation medium and without mechanical stimulation for 7 days. The expression level is expressed as mean ± SD, (n = 3) * indicates a significant difference between the different MSCs, p < 0.05. (B – G) DAB staining of Collagen 1, 2, and Aggrecan for two different cell cultures without mechanical stimulation for 3 weeks. (B – D) Normal MSCs differentiation has Collagen 1, 2, and Aggrecan matrix staining (pointed by arrows); (E – G) RohA gene-activated MSCs differentiation has obvious positive matrix staining of Collagen 1, 2, and Aggrecan (pointed by arrows). (H) Relative gene expression quantified by qPCR of MSCs differentiation without mechanical stimulation, with mechanical stimulation, or with mechanical stimulation and RhoA inhibitor C3 for 7 days. The expression level is expressed as mean ± SD, (n = 3)*indicates a significant difference compared to the cell culture without mechanical stimulation, p < 0.05. # indicates a significant difference compared to the cell culture with mechanical stimulation, p < 0.05.

Article Snippet: Diaminobenzidine (DAB, ZLI-9018, ZSGB-BIO, China) staining of the main components of annulus fibrosus collagen 1 (COL1) (1:200, ab34710, Abcam, United States), collagen 2 (COL2) (1:200, ab34712, Abcam, United States), mechanical stimulation related protein RhoA (1:100, 10749-1-AP, Proteintech, China) and ROCK1 (1:200, ab97592, Abcam, United States) were conducted according to the product instructions and viewed under a microscope (IX73, Olympus, Japan).

Techniques: Gene Expression, Expressing, Staining, Cell Culture