Horseradish peroxidase hrp conjugated goat anti mouse secondary antibody

Tissue sections were obtained were from the cerebral cortex at the level of the internal capsule and thalamus. Slides were deparaffinized at 60 °C for 40 min and then submerged in heated citrate buffer for antigen retrieval prior to immunostaining. Immunofluorescent staining was performed on PN7 tissue sections using an anti-Olig2 primary antibody (1:750; Millipore, AB9610) and a donkey anti-mouse fluorescent secondary (1–500; Invitrogen, A- 21206). Slides from tissues obtained at PN21 or 4 mon were evaluated by immunohistochemistry using anti-Myelin Basic Protein (MBP) primary antibody (1:200, Abcam, ab62631), detected with horseradish peroxidase (HRP)-conjugated goat anti-mouse secondary antibody (1–600; Biorad, 170-6516) and visualized with NovaRed (Vector). The lateral cortex is defined as the area lateral to the caudoputamen in the perirhinal or ectorhinal cortical areas. The medial cortex is defined as the cortical region superior to the cingulum. Quantification was performed from 3 to 4 individual mice in two distinct regions conducted by two individuals who were blinded to the experimental conditions. Data was then averaged from the blinded individuals and statistically analyzed.

All cell culture reagents were purchased from Corning Life Sciences. Antibodies against β-tubulin, Yy1, and RhoA were purchased from Santa Cruz Biotechnology. Antibodies against Mlc; phospho-Mlc (Ser19); total Caspase-7 and Caspase-9; cleaved Caspase-3, Caspase-7, Caspase-8, and Caspase-9; cleaved PARP; Calpain-1; Calpain-2; and H3 histone were purchased from Cell Signaling Technology. Anti-Mef2D antibody was purchased from BD Biosciences. Horseradish peroxidase (HRP)-conjugated goat anti-mouse secondary antibody was purchased from Bio-Rad, and HRP-conjugated goat anti-rabbit antibody was purchased from Cell Signaling Technology. Anti-α smooth muscle actin (α-SMA) antibody was purchased from R&D Systems. All chemicals were purchased from Sigma-Aldrich. Reagents for quantitative polymerase chain reaction (qPCR) were purchased from Life Technologies. The mRNA Isolation Kit was purchased from QIAGEN.