Proteins were separated by 10% SDS-PAGE and then transferred to PVDF membranes (Amersham, Buckinghamshire, UK). The membranes were incubated overnight at 4 °C with anti-PTH1R antibody (Sigma) and anti-GAPDH (Sigma) antibody followed by HRP-linked secondary antibodies. incubated overnight at 4 °C with anti-PTH1R antibody (Sigma) and anti-GAPDH (Sigma) antibody followed by HRP-linked secondary antibodies.
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