Figure 6
From: The Na/K-ATPase α1 and c-Src form signaling complex under native condition: A crosslinking approach
Knockout of Src kinase family reduces binding of the α1 subunit with other proteins - comparison of α1 subunit-c-Src binding pattern between SYF and SYF + c-Src cells by capillary immunoblotting: Preparation of whole cell lysate with Native-PAGE sample buffer were performed as described for BN-PAGE in the Materials and Methods. Samples (3 μg protein/sample) were electrophoresed side-by-side on the same 66–440 kDa separation module and immunoblotted with detection module. For DTT/SDS treatment, samples were heated at 60 °C for 30 min for the α1 subunit detection. Control (CTL), BMH- and DTME-crosslinked samples without DTT/SDS pretreatment of SYF cells (a) and SYF + c-Src cells (b), or with DTT/SDS pretreatment of SYF cells (c) and SYF + c-Src cells (d), and then immunoblotted for α1 subunit. Please note the differences of α1 pattern between SYF and SYF + c-Src cells. n = 3.
