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The specific marker genes with primers used for quantitative reverse transcription PCR analysis.

Journal: Acta Physiologica (Oxford, England)

Article Title: Cold‐induced fibrosis and metabolic remodeling in the turtle ( Trachemys scripta ) ventricle

doi: 10.1111/apha.70026

Figure Lengend Snippet: The specific marker genes with primers used for quantitative reverse transcription PCR analysis.

Article Snippet: Conditioned media from HepG2 cell cultures (100 ng protein/lane) and recombinant active human MMP‐2 (1 ng protein/lane, Millipore) were used as positive controls.

Techniques: Marker, Reverse Transcription

Regulation of ventricular connective tissue. (A–D) Quantitative real‐time PCR analysis of mRNA expression of (A) tissue inhibitor of matrix metalloproteinases (TIMP2), (B) collagen degrading matrix metalloproteinase‐2 (MMP2), (C) matrix metalloproteinase‐9 (MMP9), and (D) collagen I gene (COL1α2), for cold‐acclimated (5°C; striped) and control (25°C; filled) freshwater turtles ( n = 5). Values are presented as mean ± SE . Individual dots are individual turtles. Significant differences in mRNA expression were assessed by GLM and indicated by * ( p < 0.05). (E–G) Characterization of specific matrix metalloproteinase (MMP) activity in the ventricular myocardium by gelatin SDS‐PAGE zymography. (E) Coomassie R250 stained zymogram, indicating the relative molecular weights (MW, kDa) and abundances of gelatinases in ventricle extracts from cold‐acclimated (blue) and control (red) freshwater turtles ( n = 5). The positions of MMP9, proMMP‐2 and MMP2 are indicated by arrows on the left‐hand side of the gel. (F) Abundance of MMP9 and (G) abundance of MMP2 from ventricle extracts of cold‐acclimated and control freshwater turtle ( n = 5). Values are presented as mean ± SE . Individual dots are individual turtles. No significant differences in MMP abundance were found when assessed by GLM. (H, I) Endogenous matrix metalloproteinase (MMP) activity. (H) Representative fluorescent micrographs for cold‐acclimated and control ventricle imaged with a green filter to show gelatinase activity. The same sections were imaged in with a blue filter and the image imposed to show DAPI fluorescence for cold‐acclimated and control turtle tissue. (I) Semi‐quantitative analysis of endogenous MMP activity by in situ gelatinase zymography of ventricular sections for cold‐acclimated and control freshwater turtles ( n = 5). Values are presented as mean ± interquartile range and range. Individual dots are individual turtles. Significant differences in collagen coherency were assessed by GLM and indicated by * ( p < 0.05).

Journal: Acta Physiologica (Oxford, England)

Article Title: Cold‐induced fibrosis and metabolic remodeling in the turtle ( Trachemys scripta ) ventricle

doi: 10.1111/apha.70026

Figure Lengend Snippet: Regulation of ventricular connective tissue. (A–D) Quantitative real‐time PCR analysis of mRNA expression of (A) tissue inhibitor of matrix metalloproteinases (TIMP2), (B) collagen degrading matrix metalloproteinase‐2 (MMP2), (C) matrix metalloproteinase‐9 (MMP9), and (D) collagen I gene (COL1α2), for cold‐acclimated (5°C; striped) and control (25°C; filled) freshwater turtles ( n = 5). Values are presented as mean ± SE . Individual dots are individual turtles. Significant differences in mRNA expression were assessed by GLM and indicated by * ( p < 0.05). (E–G) Characterization of specific matrix metalloproteinase (MMP) activity in the ventricular myocardium by gelatin SDS‐PAGE zymography. (E) Coomassie R250 stained zymogram, indicating the relative molecular weights (MW, kDa) and abundances of gelatinases in ventricle extracts from cold‐acclimated (blue) and control (red) freshwater turtles ( n = 5). The positions of MMP9, proMMP‐2 and MMP2 are indicated by arrows on the left‐hand side of the gel. (F) Abundance of MMP9 and (G) abundance of MMP2 from ventricle extracts of cold‐acclimated and control freshwater turtle ( n = 5). Values are presented as mean ± SE . Individual dots are individual turtles. No significant differences in MMP abundance were found when assessed by GLM. (H, I) Endogenous matrix metalloproteinase (MMP) activity. (H) Representative fluorescent micrographs for cold‐acclimated and control ventricle imaged with a green filter to show gelatinase activity. The same sections were imaged in with a blue filter and the image imposed to show DAPI fluorescence for cold‐acclimated and control turtle tissue. (I) Semi‐quantitative analysis of endogenous MMP activity by in situ gelatinase zymography of ventricular sections for cold‐acclimated and control freshwater turtles ( n = 5). Values are presented as mean ± interquartile range and range. Individual dots are individual turtles. Significant differences in collagen coherency were assessed by GLM and indicated by * ( p < 0.05).

Article Snippet: Conditioned media from HepG2 cell cultures (100 ng protein/lane) and recombinant active human MMP‐2 (1 ng protein/lane, Millipore) were used as positive controls.

Techniques: Real-time Polymerase Chain Reaction, Expressing, Control, Activity Assay, SDS Page, Zymography, Staining, Fluorescence, In Situ