Heartattack study could change the game in regenerativemedicine sbpdiscovery NatureComms
. In brief, HDFs were cultured and harvested as described above, resuspended in FCM, added to 384-well plates at 3 × 10cells/well, and transfected with siRNAs. The next day , 0.25 µL of F-ABM lentiviral mix was added to each well, and the cells were incubated for an additional 24 h. The following day , ABM expression was induced by the addition of 2 μg/mL Dox diluted in FCM.
. In brief, HDFs were cultured and harvested as described above, resuspended in FCM, added to 384-well plates at 1 × 10cells/well, and transfected with siRNAs. Cells settled to the bottom of the well for 5 min at room temperature and then 0.25 µL of F-OKMS lentiviral mix was added to each well, and the cells were incubated for 24 h. The following day , OKMS expression was induced by the addition of 2 μg/mL Dox diluted in FCM.
Mouse heart sections were fixed with 4% PFA for 10 min. Sections were permeabilized and blocked in 20% goat serum and 0.3% Triton X-100 in PBS for 1 h. Incubation with the primary antibody was performed overnight at 4 °C. Sections were washed with PBS. Then, the secondary antibody was left for 1 h at room temperature. Nuclei were counterstained with DAPI for 10 min. Slides were mounted with Fluoromount-G .
The primary antibodies used were: rabbit anti-ATF7IP ; rabbit anti-ATF7IP ; rabbit anti-JUNB ; rabbit anti-SP7/OSTERIX ; mouse anti-ZNF207 ; rabbit anti-TAGLN ; guinea pig polyclonal anti-Vimentin ; mouse anti-VIMENTIN ; goat polyclonal anti-TAGLN ; rabbit polyclonal anti-TNNT2 ; mouse anti-ACTN2 ; goat anti-PECAM1 ; rabbit anti-MAP2 ; and mouse anti-TUJ1 ; rabbit anti-NANOG ; rabbit anti-Collagen I .
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