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Blots were washed three times with 1% (w/v) non-fat dry milk TTBS and incubated with the secondary antibody (rabbit for Id2 and mouse for tubulin) diluted in 1% non-fat dry milk TTBS conjugated to horseradish peroxidase (Santa Cruz Biotechnology, CA, USA) for 60 min at room temperature

Experimental data indicate that C-peptide can bind to cell surface receptors, likely involving G-protein, coupled mechanisms, and influence downstream pathways related to nitric oxide production, calcium signaling, and kinase activation