Rabbit anti prdm16

Mouse embryos and neonates were collected, fixed in 4% PFA, embedded in OCT, and frozen in dry iced hexane. Frozen sections were blocked with 5% goat serum, stained with mouse anti-TNNT2 (Thermo Scientific), mouse anti-sarcomeric alpha-actinin (Sigma Aldrich), rabbit anti-TNNI3 (Santa Cruz), rabbit anti-TGFβ1 (Abcam), rabbit anti-Ki67 (Thermo Scientific), rabbit anti-phospho-Histone H3 (Cell Signaling Technology), and Hoechst 33342, and visualized using Alexa Fluor-conjugated secondary antibodies (Life Technologies). Image acquisition was performed on a Zeiss LSM510 Meta inverted confocal microscope (Carl Zeiss) or an Eclipse 80i fluorescent microscope (Nikon). Transgenes were detected by PCR from yolk sac DNA of embryos or tail DNA of neonatal pups. For human heart tissues, formalin-fixed paraffin embedded tissue-sections were rehydrated and autoclaved with Citrate Buffer, pH 6.0 (Sigma-Aldrich) and stained with rabbit anti-phospho-Smad2 (EMD Millipore), rabbit anti-PRDM16 (Abcam), mouse anti-sarcomeric alpha-actinin (Sigma Aldrich) and Hoechst 33342, and visualized using Alexa Fluor-conjugated secondary antibodies (Life Technologies). Image acquisition was performed on an Eclipse 80i fluorescent microscope (Nikon). A detailed list of antibodies used is shown in Supplementary Table 10.