Journal: Oncotarget

Article Title: Na v 1.5 regulates breast tumor growth and metastatic dissemination in vivo

doi:

Figure Lengend Snippet: A. Representative images of non-cancer breast tissue and B. breast tumor tissue in which Na v 1.5 was (i) absent, (ii) weakly positive, and (iii) strongly positive. C. Absence of immunoreactivity in a “strongly positive” tumor stained with anti-Na v 1.5 antibody preincubated with immunizing peptide. D. Adjacent section from the same tumor in (Biii) stained with anti-β1 antibody . Scale bar, 100 μm. Insets, higher magnification images of tumor cells, scale bar, 50 μm. E. Mean Allred score for matched normal breast and tumor samples. Data are mean + SEM ( n = 36 matched cases). *** P < 0.001. F. Na v 1.5 vs. β1 IHC score in adjacent sections from matched tumor samples. Pearson r = 0.66 ( P < 0.001). Solid line, linear regression; broken lines 95% confidence intervals. G. Western blot of Na v 1.5 expression across a panel of BCa cell lines. Loading control = heat shock protein 90 (HSP90). Positive control = HEK293 cells stably expressing Na v 1.5 .

Article Snippet: Immunohistochemistry was performed using the EnVision+ System-HRP kit (Dako) and anti-Na v 1.5 antibody (1:100; Alomone) [ , ].

Techniques: Staining, Western Blot, Expressing, Positive Control, Stable Transfection

Journal: Oncotarget

Article Title: Na v 1.5 regulates breast tumor growth and metastatic dissemination in vivo

doi:

Figure Lengend Snippet: A. Western blot of Na v 1.5 in MDA-MB-231 cells stably expressing control shRNA, or shRNA1, or shRNA2. Loading control = heat shock protein 90 (HSP90). B. Typical in vitro whole-cell recordings from MDA-MB-231 cells stably expressing control shRNA, or shRNA1, or shRNA2. Recordings were taken after depolarization to voltages in the range −80 to +30 mV in 5 mV steps following a 250 ms prepulse at −120 mV. C. Peak Na + current density and D. current-voltage relationship of MDA-MB-231 cells stably expressing control shRNA, or shRNA1, or shRNA2 ( n = 8/each). E. Representative bioluminescent images of mice bearing tumors containing MDA-MB-231 cells stably expressing control shRNA (“Control”) and MDA-MB-231 cells expressing shRNA1 (“shRNA”), 4 weeks after implantation. F. Bioluminescence measured from primary tumors on the indicated days post-implantation ( n ≥ 13). G. Calculated volume derived from caliper measurement of primary tumors over the same period ( n ≥ 13). Data are mean ± SEM; ** P < 0.01; *** P < 0.001.

Article Snippet: Immunohistochemistry was performed using the EnVision+ System-HRP kit (Dako) and anti-Na v 1.5 antibody (1:100; Alomone) [ , ].

Techniques: Western Blot, Stable Transfection, Expressing, shRNA, In Vitro, Derivative Assay

Journal: Oncotarget

Article Title: Na v 1.5 regulates breast tumor growth and metastatic dissemination in vivo

doi:

Figure Lengend Snippet: Na + influx carried by Na v 1.5 allosterically regulates NHE1 to increase H + efflux, lowering extracellular pH and enhancing cathepsin protease activity [ , ]. Na v 1.5 also increases src activity and cortactin phosphorylation, resulting in cytoskeletal changes, invadopodia formation and acquisition of an elongate invasive morphology . The β1 subunit enhances process outgrowth via a trans -homophilic adhesion mechanism that requires fyn kinase and Na + current . Na v 1.5 positively regulates CD44 protein expression, which may in turn enhance CD44-mediated src activation and increase cortactin expression/activity [ – ]. Na v 1.5 expression is maintained by a positive feedback mechanism via protein kinase A (PKA) [ , ]. Additional potential partners in this signaling network have been omitted for clarity, and are reviewed in detail elsewhere [ , ]. Figure was produced using ScienceSlides software.

Article Snippet: Immunohistochemistry was performed using the EnVision+ System-HRP kit (Dako) and anti-Na v 1.5 antibody (1:100; Alomone) [ , ].

Techniques: Activity Assay, Expressing, Activation Assay, Produced, Software