4.4. Western Blot

H⁺, K⁺-ATPase subunits detection was performed as described previously [15]. For signaling pathway analysis, PANC-1 cell lysate was boiled 3 min at 95 °C. Primary antibodies against gastric HKα1 (Abcam {"type":"entrez-protein","attrs":{"text":"EPR12251","term_id":"523377848","term_text":"EPR12251"}}EPR12251, 1:1000), non-gastric HKα2 (C384-M79, 1:1000, kindly donated by J. J. H. H. M. De Pont and H. G. P. Swarts) [56], HKβ (Sigma A274, RRID:AB_258027, 1:1000), β-Actin (Santa Cruz Sc-47778, 1:1000, RRID: AB_626632), pSTAT3 (Cell Signaling #9145, 1:1000, RRID: AB_2491009), STAT3 (Cell Signaling #9139, 1:1000, RRID: AB_331757) and Cyclin D1 (Millipore #06-137, 1:500, RRID: AB_2070695) were used.