Journal: PLoS ONE

Article Title: Krüppel-Like Factor 2 Is Required for Normal Mouse Cardiac Development

doi: 10.1371/journal.pone.0054891

Figure Lengend Snippet: Immunohistochemical staining (n = 2) was performed using an endothelial cell specific mouse PECAM (CD31) antibody. A goat anti-mouse secondary antibody conjugated with HRP was used, and reacted with diaminobenzidine (DAB) for detection. Brown coloration of cells indicates a CD31 positive cell type. ( A ) The cells lining the AV canal in FVB/N WT are CD31 positive. ( B ) Higher magnification of WT AV region. ( C ) and ( E ) Cells accumulated in the AV canal in two different FVB/N KLF2−/− hearts are CD31 positive, indicating that they are endothelial cells. ( D ) and ( F ) Higher magnifications of KLF2−/− AV regions show accumulation of stained cells. Asterisks* indicate AV endothelial cushion region. A, C and E are 100X magnification; B, D and F are 200X. ( G ) qRT-PCR shows a 2-fold increase in expression of PECAM mRNA in FVB/N KLF2−/− AV canals compared to FVB/N WT (p = 0.002, n = 7). The amount of PECAM mRNA in WT was designated 100%.

Article Snippet: The embryos were incubated with secondary antibody, biotinylated Anti-Rat IgG (1∶500) (Abcam) and BD Pharmingen Streptavidin-Horseradish Peroxidase (Sav-HRP) and BD Pharmingen 3,3′diaminobenzidine (DAB) chromogen in H 2 O 2 buffer and fixed in 4% PFA.

Techniques: Immunohistochemical staining, Staining, Quantitative RT-PCR, Expressing