Human/Mouse Myocardin Antibody

Detection of Human Myocardin by Western Blot. Western blot shows lysates of MCF-7 human breast cancer cell line, Raji human Burkitt's lymphoma cell line, HeLa human cervical epithelial carcinoma cell line, and C2C12 mouse myoblast cell line. PVDF membrane was probed with 1 µg/mL of Human Myocardin Monoclonal Antibody (Catalog # MAB4028) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Myocardin at approximately 105 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Mouse Myocardin by Western Blot Laminin-111 but not laminin-alpha 4 blocking antibody affects pericyte differentiation. (a) Immunoblots show that laminin-111 blockage (Ln Ab) significantly enhances the expression of PDGFR beta, SMA, and SM22-alpha, but not myocardin in pericytes. Full blots of these proteins are shown in Supplementary Figure 14b. Rabbit IgG treated cells were used as a Ctr. All bands were normalized to actin (n=5–6). (b) Immunoblots show that laminin-alpha 4 blockage (Anti-Ln alpha 4) does not change the expression of PDGFR beta, SMA, SM22-alpha, or myocardinin in pericytes. Full blots of these proteins are shown in Supplementary Figure 14c. Rabbit IgG treated cells were used as a Ctr. All bands were normalized to actin (n=3). Data are shown as mean ± sd. *p<0.05 versus the Ctrs by student’s t-test. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/24583950), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse Myocardin by Western Blot Astrocytic laminin mediates pericyte differentiation via integrin alpha 2. (a) Immunoblots show that integrin alpha 2 blockage (ITGA2) but not integrin beta 1 blockage significantly increases the expression of PDGFR beta, SMA, and SM22-alpha, but not myocardin in pericytes. Full blots of these proteins are shown in Supplementary Figure 14d. Rabbit IgG treated cells were used as a Ctr. All bands were normalized to actin (n=6). (b) Schematic diagram of shRNA designed to target ITGA2 mRNA. (c) Immunoblot analysis shows that all three ITGA2-specific shRNAs (#1–3) dramatically reduce ITGA2 at protein level and ITGA2-specific shRNA-3 (#1) is the most efficient one. Full blots of ITGA2 and actin are shown in Supplementary Figure 14e. Scramble shRNA was used as a Ctr. (d) Immunoblot analysis shows that transduction of pericytes with lenti-shRNA-1 (#1) significantly enhances the expression of PDGFR beta, SMA, and SM22-alpha, but does not affect myocardin level. Full blots of these proteins are shown in Supplementary Figure 14f. Scramble shRNA was used as a Ctr. All bands were normalized to actin (n=4–5). Data are shown as mean ± sd. *p<0.05 versus the Ctrs by student’s t-test. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/24583950), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse Myocardin by Western Blot Laminin-111 but not laminin-alpha 4 blocking antibody affects pericyte differentiation. (a) Immunoblots show that laminin-111 blockage (Ln Ab) significantly enhances the expression of PDGFR beta, SMA, and SM22-alpha, but not myocardin in pericytes. Full blots of these proteins are shown in Supplementary Figure 14b. Rabbit IgG treated cells were used as a Ctr. All bands were normalized to actin (n=5–6). (b) Immunoblots show that laminin-alpha 4 blockage (Anti-Ln alpha 4) does not change the expression of PDGFR beta, SMA, SM22-alpha, or myocardinin in pericytes. Full blots of these proteins are shown in Supplementary Figure 14c. Rabbit IgG treated cells were used as a Ctr. All bands were normalized to actin (n=3). Data are shown as mean ± sd. *p<0.05 versus the Ctrs by student’s t-test. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/24583950), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse Myocardin by Western Blot Astrocytic laminin mediates pericyte differentiation via integrin alpha 2. (a) Immunoblots show that integrin alpha 2 blockage (ITGA2) but not integrin beta 1 blockage significantly increases the expression of PDGFR beta, SMA, and SM22-alpha, but not myocardin in pericytes. Full blots of these proteins are shown in Supplementary Figure 14d. Rabbit IgG treated cells were used as a Ctr. All bands were normalized to actin (n=6). (b) Schematic diagram of shRNA designed to target ITGA2 mRNA. (c) Immunoblot analysis shows that all three ITGA2-specific shRNAs (#1–3) dramatically reduce ITGA2 at protein level and ITGA2-specific shRNA-3 (#1) is the most efficient one. Full blots of ITGA2 and actin are shown in Supplementary Figure 14e. Scramble shRNA was used as a Ctr. (d) Immunoblot analysis shows that transduction of pericytes with lenti-shRNA-1 (#1) significantly enhances the expression of PDGFR beta, SMA, and SM22-alpha, but does not affect myocardin level. Full blots of these proteins are shown in Supplementary Figure 14f. Scramble shRNA was used as a Ctr. All bands were normalized to actin (n=4–5). Data are shown as mean ± sd. *p<0.05 versus the Ctrs by student’s t-test. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/24583950), licensed under a CC-BY license. Not internally tested by R&D Systems.