Fig. 6

PMCA expression was significantly induced under metabolic or anti-cancer drug stress. Immunohistochemical analysis of PMCA protein levels in paraffin-embedded xenograft tumor tissues with metabolic and drug-resistant (patient-derived) cancer cells. A S-231 (top, PMCA1) and S-MCF-7 (bottom, PMCA2) were examined at 400 × magnification; scale bar: 80 μm. B S-231 (left) and S-MCF-7 (right), image analysis software was used to quantify the immunostained target proteins. Each assay was performed in triplicate and representative images are displayed. *P < 0.05 and **P < 0.01 vs. control. C Immunoblot analysis for CHOP (ER stress marker) expression after treatment of 2DG (2-deoxy-glucose, glucose deprivation-induced metabolic stress inducer) alone or with caloxin or candidate 13 (PMCA inhibitor) in a mouse xenograft model of selected cells (glucose deprivation-induced metabolic stress-resistant cancer cells). D Immunohistochemical analysis of PMCA protein levels in paraffin-embedded xenograft tumor tissues with patient-derived drug-resistant cancer cells after treatment of oxaliplatin or sorafenib alone or with caloxin or candidate 13 (PMCA inhibitor). YUMC-C1 (top), YUMC-C2 (middle), and YUMC-P1 (bottom). E Each assay was performed in triplicate; representative images were generated with MetaMorph 4.6. YUMC-C1 (left), YUMC-C2 (middle), and YUMC-P1 (right). *P < 0.05 and **P < 0.01 vs. control. F Immunoblot analysis for CHOP (ER stress marker) expression after treatment with oxaliplatin or sorafenib alone or with caloxin or candidate 13 in a mouse xenograft model with patient-derived drug-resistant cancer cells. G, H Immunohistochemical analysis and image analysis of PMCA protein levels in paraffin-embedded patient tumor tissues with neoadjuvant resistance to ER +/− (estrogen receptor positive or negative). **P < 0.01 vs. neoadjuvant resistance/ER+ or ER−. Each assay was performed in triplicate, and representative images are displayed