Orai3

Cell lysates were harvested in RIPA buffer (150 mM NaCl, 1 mM EGTA, 50 mM Tris at pH 7.4, 10% glycerol, 1% Triton X-100, 1% sodium deoxycholate, 0.1% SDS, and Complete^TM), and subsequently analyzed by Western blotting using antibodies against STIM1, FAK, paxillin, E-cadherin, ZO-1, fibronectin (BD Biosciences, San Jose, CA), Orai1, Orai3 (Prosci, Poway, CA), Orai2, phospho-Tyr397-FAK (Enzo, Farmingdale, NY), TRPC1 (Proteintec, Rosemont, IL), vinculin, vimentin, N-cadherin (Santa Cruz, Santa Cruz, CA), talin (abcam, Cambridge, UK), phospho-Tyr18-paxillin (Invitrogen, San Diego, CA), STIM2 (Cell Signaling, Danvers, MA), and β-actin (Sigma-Aldrich, Saint Louis, MO). The immunocomplexes were then detected with horseradish peroxidase-conjugated IgG (Jackson ImmunoResearch Laboratories, West Grove, PA), and the reaction was developed using an ECL detection kit (Amersham, Piscataway, NJ) under an ImageQuant LAS 4000 system (GE Healthcare Life Sciences, Pittsburgh, PA).

Cell lysates were harvested in RIPA buffer (150 mM NaCl, 1 mM EGTA, 50 mM Tris at pH 7.4, 10% glycerol, 1% Triton X-100, 1% sodium deoxycholate, 0.1% SDS, and Complete^TM), and the lysates were analyzed by Western blotting using antibodies against Bcl-2 (DAKO, Grostrup, Denmark), Bax, Bak, calnexin, SERCA2, β-actin (Santa Cruz Biotechnology, Santa Cruz, CA), porin (Molecular Probes, Eugene, OR), caspase-8, caspase-9, caspase-12, SERCA3, pSer70-Bcl-2, pThr167-Bax (Cell Signaling Technology, Beverly, MA), Grp78, STIM1, STIM2, IP3R3 (BD, Franklin Lakes, NJ), Orai1, Orai2, Orai3, and TRPC1 (ProSci, Poway, CA).

Cell lysates were harvested in radioimmunoprecipitation assay (RIPA) buffer (150 mM NaCl, 1 mM EGTA, 50 mM Tris at pH 7.4, 10% glycerol, 1% Triton X-100, 1% sodium deoxycholate, 0.1% SDS, and CompleteTM), and the lysates were analyzed by Western blotting using antibodies against STIM1 (BD, Franklin Lakes, NJ, USA), Orai1, Orai3 (ProSci, Poway, CA, USA), Orai2 (Enzo, Farmingdale, NY, USA), TRPC1 (Proteintech, Rosemont, IL, USA), PDGFRα, JNK, phospho-EGFR, EGFR (Santa Cruz, Santa Cruz, CA, USA), phospho-PDGFRβ, PDGRβ (Abnova, San Francisco, CA, USA), STIM2, phospho-PLCγ, PLCγ, phospho-Akt, Akt, phospho-JNK, phospho-ERK, ERK, phospho-STAT3, STAT3, phospho-CREB, CREB (Cell Signaling, Beverly, MA, USA), and β-actin (Sigma, Saint Louis, MO, USA). The immune complexes were then detected with horseradish peroxidase-conjugated IgG (Jackson ImmunoResearch Laboratories, West Grove, PA, USA), and the reaction was developed using an enhanced chemiluminescence (ECL) detection kit under an ImageQuant LAS 4000 system (GE Healthcare Life Sciences, Pittsburgh, PA, USA).