Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Amiloride sensitizes prostate cancer cells to the reversible tyrosine kinase inhibitor lapatinib by modulating Erbb3 subcellular localization

doi: 10.1007/s00018-024-05540-5

Figure Lengend Snippet: Amiloride promoted ErbB3 translocation from the nucleus to the cytoplasm and the plasma membrane in HSPC cells ( A ) Hormone-sensitive LNCaP cells were treated with varying concentrations of amiloride for 72 h before being lysed, fractionated and analyzed by immunoblot. ( B ) Immunofluorescence microscopy (IF) in LNCaP cells treated with DMSO or 75 µM or and probed with IF specific C-terminal ErbB3 antibodies or ( C ) 10µM or 25µM amiloride and probed with IF specific N-terminal ErbB3 antibodies for 72 h (scale bars = 30 μm). Note that vehicle treated LNCaP cells expressed nuclear ErbB3 (red) whereas amiloride-treated cells had significantly decreased ErbB3 expression in the nucleus (hollowed out). Location of nuclei are identified by blue DAPI staining. Plasma membrane localization of ErbB3 at cell-cell junction was also noted in amiloride-treated but not in vehicle treated cells. Note that both N- and C-terminal ErbB3 antibodies demonstrate lighter nuclear staining in amiloride-treated cells. ( D ) Cells were subjected to viability assays using the stated concentrations of amiloride. p-values are calculated with respect to DMSO

Article Snippet: After washing thrice with cold PBST, coverslips were inverted and mounted onto uncharged glass slides with UltraCruz Hardset Mounting Medium plus DAPI (SantaCruz BioTech, Dallas, TX).

Techniques: Translocation Assay, Clinical Proteomics, Membrane, Western Blot, Immunofluorescence, Microscopy, Expressing, Staining