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1. ECM:PBS coating (1:100) + cylinder placement. Overnight @RT
2. GF/DF placement inside cylinder. Overnight @4°C
3. Removal of cylinders
4. Media and cells placement
5. Addition of BrdU 2 hours prior to fixation
6. Cells fixing after 48 hours
7. Indirect immunofluorescence (BrdU for proliferation, eMHC for differentiation, nuclear dye Hoechst)
