CTGF/CCN2 Antibody

Immunohistochemistry: CTGF/CCN2 Antibody [NB100-724] -

Immunohistochemistry: CTGF/CCN2 Antibody [NB100-724] - GFAP expression & synthesis in the ON & ONH in a murine glaucoma model. (A) GFAP immunoreactivity in the glial lamina of 1-month (green, upper panel) & 2 months (green, middle & lower panel) old betaB1-CTGF1 mice compared to WT controls. Immunoreactivity of GFAP not altered in 1-month old TG & WT animals (green, upper panel). 2 month old betaB1-CTGF1 mice showed an increased GFAP immunoreactivity compared to WT control (green, middle panel). Lower panel shows a magnified detail of the middle panel. Nuclei are stained w/ Dapi (blue). (B) Schmatic illustration of the optic nerve. A depicts region in the glial lamina where cross sections obtained. ONH (unmyelinated part) & ON (myelinated part) used for molecular analysis. (C) Quantification of immunohistochemical staining of GFAP in the glial lamina is not altered in 1 month old TG & WT animals (WT: n = 5; TG: n = 4). GFAP immunoreactivity is increased in 2 month old TG animals compared to WT littermates (WT: n = 10, TG: n = 13; *p = 0.039; two-tailed t-test compared to oretical mean of 1 (normalized control)). Mean value of WT animals (control) set at 1. (D) RT-PCR analyses revealed no alteration in the Gfap mRNA expression in the ON of 1-month (WT: n = 7; TG: n = 5) & 2 month old betaB1-CTGF1 mice (WT: n = 15; TG: n = 16) compared to WT controls. In the ONH the Gfap mRNA expression is increased in the 2 month old TG compared to WT animals (WT: n = 6, TG: n = 5; *p = 0.04). The mRNA expression of Gfap is not altered in 1 month old animals (WT: n = 6, TG: n = 5). Mean value of WT animals (control) set at 1. RPL32 used to normalize mRNA expression. Data represented as mean ± SEM. PRL, Prelaminar region; PSL, Postlaminar region; ON, optic nerve; ONH, optic nerve head. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/35493079), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: CTGF/CCN2 Antibody [NB100-724] -

Immunohistochemistry: CTGF/CCN2 Antibody [NB100-724] - GFAP expression & synthesis in the ON & ONH in a murine glaucoma model. (A) GFAP immunoreactivity in the glial lamina of 1-month (green, upper panel) & 2 months (green, middle & lower panel) old betaB1-CTGF1 mice compared to WT controls. Immunoreactivity of GFAP not altered in 1-month old TG & WT animals (green, upper panel). 2 month old betaB1-CTGF1 mice showed an increased GFAP immunoreactivity compared to WT control (green, middle panel). Lower panel shows a magnified detail of the middle panel. Nuclei are stained w/ Dapi (blue). (B) Schmatic illustration of the optic nerve. A depicts region in the glial lamina where cross sections obtained. ONH (unmyelinated part) & ON (myelinated part) used for molecular analysis. (C) Quantification of immunohistochemical staining of GFAP in the glial lamina is not altered in 1 month old TG & WT animals (WT: n = 5; TG: n = 4). GFAP immunoreactivity is increased in 2 month old TG animals compared to WT littermates (WT: n = 10, TG: n = 13; *p = 0.039; two-tailed t-test compared to oretical mean of 1 (normalized control)). Mean value of WT animals (control) set at 1. (D) RT-PCR analyses revealed no alteration in the Gfap mRNA expression in the ON of 1-month (WT: n = 7; TG: n = 5) & 2 month old betaB1-CTGF1 mice (WT: n = 15; TG: n = 16) compared to WT controls. In the ONH the Gfap mRNA expression is increased in the 2 month old TG compared to WT animals (WT: n = 6, TG: n = 5; *p = 0.04). The mRNA expression of Gfap is not altered in 1 month old animals (WT: n = 6, TG: n = 5). Mean value of WT animals (control) set at 1. RPL32 used to normalize mRNA expression. Data represented as mean ± SEM. PRL, Prelaminar region; PSL, Postlaminar region; ON, optic nerve; ONH, optic nerve head. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/35493079), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: CTGF/CCN2 Antibody [NB100-724] -

Immunocytochemistry/ Immunofluorescence: CTGF/CCN2 Antibody [NB100-724] - Analyses of ECM components in murine ON astrocytes following treatment w/ TGF-beta 2 & CCN2/CTGF. (A) Immunocytochemical staining against fibronectin (green, upper panel) showed a markedly increase following the treatment w/ 1 ng/ml TGF-beta 2 or 50 ng/ml CCN2/CTGF. Immunocytochemical staining against tropoelastin (green, lower panel) showed a pronounced increase after treatment w/ 1 ng/ml TGF-beta 2 or 50 ng/ml CCN2/CTGF. Nuclei stained w/ Dapi (blue). (B) Real-time RT-PCR analyses shows an intense upregulation of collagen 3 alpha1, tropoelastin & fibronectin mRNA in murine ON astrocytes after treatment w/ 1 ng/ml TGF-beta 2, 50 ng/ml CCN2/CTGF or 100 ng/ml CCN2/CTGF for 24 h compared to untreated control cells (collagen 3 alpha1: n = 3; TGF-beta 2 ***p = 0.0000004, 50 ngCTGF **p = 0.005, 100ngCTGF **p = 0.008; fibronectin: n = 6, TGF-beta 2 *p = 0.00004, 50 ng CTGF *p = 0.016, 100 ngCTGF *p = 0.05; elastin: n = 5, TGF-beta 2 *p = 0.026, 50 ng CTGF *0.04, 100 ng CTGF *p = 0.03). mRNA expression normalized to Gnb2l & mean value of control cells set at 1. (C) Western Blot analysis show an increase in protein synthesis of collagen 3 alpha1, elastin & fibronectin in murine ON astrocytes after treatment w/ 1 ng/ml TGF-beta 2, 50 ng/ml CCN2/CTGF or 100 ng/ml CCN2/CTGF for 24 h compared to untreated control cells (collagen 3 alpha1: n = 3, TGF-beta 2 *p = 0.03, 100 ngCTGF **p = 0.009; fibronectin: n = 5, **p = 0.005, 50 ngCTGF **p = 0.006, 100 ng CTGF *p = 0.02; tropoelastin: n = 5, TGF-beta 2 ***p = 0.0004, 50 ngCTGF **p = 0.002, 100 ngCTGF ***p = 0.00001). GAPDH & alpha-tubulin used to normalize protein synthesis & mean value of control cells set at 1. Right panel shows representative Western Blots for all three proteins. Data represented as mean ± SD. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/35493079), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: CTGF/CCN2 Antibody [NB100-724] -

Immunocytochemistry/ Immunofluorescence: CTGF/CCN2 Antibody [NB100-724] - Increasing substratum stiffness causes alterations in murine ON astrocyte reactivity, actin cytoskeleton & CCN2/CTGF level. (A) Filamentous actin labeled by phalloidin (red, upper panel) is increased when murine ON astrocytes were cultured on substrates with higher stiffness (30 kPa, 60 kPa) compared to softer control (10 kPa). Increasing substratum stiffness leads to an enhanced formation of actin stress fibers. Immunoreactivity of CCN2/CTGF (green, middle panel) is markedly increased when murine ON astrocytes were grown on substrates with higher stiffness (30, 60 kPa) compared to softer control (10 kPa). Murine ON astrocytes show an enhanced GFAP protein synthesis (green, lower panel) when cultured on 30 or 60 kPa compared to 10 kPa. Nuclei were stained with Dapi (blue). (B) CCN2/CTGF protein synthesis is significantly increased in murine ON astrocytes grown on 60 kPa compared to 10 kPa (10 kPa: n = 5; 30 kPa: n = 5; 60 kPa: n = 5; *p = 0.04; unpaired two-tailed t-test). (C) Protein synthesis of GFAP (10 kPa: n = 6; 30 kPa: n = 6, **p = 0.005; 60 kPa: n = 4, *p = 0.04; unpaired two-tailed t-test) show an enhanced reactivity of murine ON astrocytes cultured on 30 kPa or 60 kPa compared to 10 kPa control. (D) Vimentin protein synthesis is significantly increased in murine ON astrocytes cultured on 60 kPa compared to 10 kPa (10 kPa: n = 4; 30 kPa: n = 4; 60 kPa: n = 3 ***p = 0.00002). Protein synthesis was normalized to alpha-tubulin & mean value of control (10 kPa) was set at 1. Data represented as mean ± SEM. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/35493079), licensed under a CC-BY license. Not internally tested by Novus Biologicals.