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C2C12 cells were grown in 20% foetal calf serum (FCS) containing DMEM medium and were differentiated for most experiments up to six days in 2% horse serum (HS) containing DMEM medium. Adult mouse primary myoblasts were isolated from C57BL/6 wild type 3–4 week-old mice and plated on matrigel-coated dishes. The primary myoblasts were grown in IMDM GLUTAMAX-I medium with 20% FCS and were differentiated in the same medium with 2% HS. The siRNA transfection experiments were performed as per the Lipofectamine RNAiMAX manufacturer’s protocol and cells were harvested at indicated time points of differentiation after the siRNA transfection. ON-TARGET-plus SMARTpool siRNAs for Tead1, Tead2 and Tead4 knockdown and non-targeting siRNA were purchased from Dharmacon Inc. (Chicago, Il., USA). The siRNA experiments were performed at least in triplicates. Phase contrast images were taken at 4x magnification using the EVOS digital microscope |
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Protocol tips |
C2C12 cells were grown in 20% foetal calf serum (FCS) containing DMEM medium and were differentiated for most experiments up to six days in 2% horse serum (HS) containing DMEM medium. Adult mouse primary myoblasts were isolated from C57BL/6 wild type 3–4 week-old mice and plated on matrigel-coated dishes. The primary myoblasts were grown in IMDM GLUTAMAX-I medium with 20% FCS and were differentiated in the same medium with 2% HS. The siRNA transfection experiments were performed as per the Lipofectamine RNAiMAX manufacturer’s protocol and cells were harvested at indicated time points of differentiation after the siRNA transfection. ON-TARGET-plus SMARTpool siRNAs for Tead1, Tead2 and Tead4 knockdown and non-targeting siRNA were purchased from Dharmacon Inc. (Chicago, Il., USA). The siRNA experiments were performed at least in triplicates. Phase contrast images were taken at 4x magnification using the EVOS digital microscope |
Publication protocol
C2C12 cells were grown in 20% foetal calf serum (FCS) containing DMEM medium and were differentiated for most experiments up to six days in 2% horse serum (HS) containing DMEM medium. Adult mouse primary myoblasts were isolated from C57BL/6 wild type 3–4 week-old mice and plated on matrigel-coated dishes. The primary myoblasts were grown in IMDM GLUTAMAX-I medium with 20% FCS and were differentiated in the same medium with 2% HS. The siRNA transfection experiments were performed as per the Lipofectamine RNAiMAX manufacturer’s protocol and cells were harvested at indicated time points of differentiation after the siRNA transfection. ON-TARGET-plus SMARTpool siRNAs for Tead1, Tead2 and Tead4 knockdown and non-targeting siRNA were purchased from Dharmacon Inc. (Chicago, Il., USA). The siRNA experiments were performed at least in triplicates. Phase contrast images were taken at 4x magnification using the EVOS digital microscope
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