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a combination of 1mM IBMX, 1 M dexamethasone, and 10 g/ml of insulin in media were added to fully confluent 3T3-L1 fibroblasts for three days (days 03) to initiate differentiation

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Samples were then washed with PBS as described previously and incubated with goat anti-rabbit secondary antibody (1:500 dilution, Beyotime) at room temperature for 1 h in darkness