Anti n cadherin 610921

Gold (III) chloride (520918,), Sodium Citrate (C8532), gemcitabine (G6423), and Thiazolyl Blue Tetrazolium Bromide (MTT) (M6494) were purchased from Sigma (St. Louis, MO, USA). Matrigel (CACB354234) and transwell (89235-020) were bought from VWR (Radnor, PA, USA). Alexa Fluor 488-conjugated secondary antibody (R37114) and Alexa Fluor 568-Phalloidin (A12380) were purchased from Life technology (Waltham, MA, USA). The RNA extraction kit is from ZYMO research (R1055, Irvine, CA, US). cDNA transcription kit (1708891) and the qPCR kit (1708882, iQ SYBR Green Supermix) were from Bio-Rad (Hercules, CA, USA).
Anti-E-cadherin (610182) and anti-N-cadherin (610921) were from BD Bioscience ( San Jose, CA, USA); anti-Vimentin (5741), anti-p38(8690), anti-phospho p38(9211), anti-p42/44 (9102), and anti-phospho p42/44(4370) were from Cell Signaling (Danvers, Massachusetts, US); anti-GAPDH(G9545, Sigma, St. Louis, MO, USA), anti-Tubulin(ab18207, Abcam, Cambridge, MA), DAPI( H-1500, Vector Laboratories, Burlingame, CA, USA).

Thin paraffin sections of tissue slices were stained with hematoxylin–eosin using standard techniques. Immunohistochemistry was done on whole-mount heart slices that had been fixed in 4% paraformaldehyde for at least 24 h. The tissues were equilibrated with a graded series of 4, 15 and 30% sucrose in PBS, and were permeabilized with 1% Triton X-100 overnight. After blocking (3% BSA in PBS, 12 h), samples were incubated sequentially with primary antibodies (anti-α-actinin, A7811, anti-connexin-43, C6219, anti-α-smooth muscle actin, A5228, all Sigma-Aldrich, anti-vimentin, AB92547, Abcam, anti-N-cadherin, #610921, BD-Biosciences, all 1:100), and secondary antibodies (anti-rabbit-Alexa488, A21441, anti-mouse-Alexa546, A11030, both ThermoFisher, 1:100, combined with DNA-stain 1 µM TO-PRO-3) for 1 day each. Washing steps between all incubations used citrate-buffer (150 mM NaCl, 15 mM Na₃-citrate, pH 7.2) supplemented with 1% BSA, 0.05% Triton X-100 and 3 mM NaN₃. Slices were mounted in VectaMount AQ (Vector Laboratories). Confocal microscopy was performed at the bioimaging core facility of the Biomedical Center using an inverted Leica SP8X WLL microscope.