Cyclin d1 ab16663

HT-29 cells (1 × 10⁷) were treated with PCA (362 and 724 μM) for 48 h, and then collected for western blot analysis by a previously described method [38 (link)]. Total protein was measured by the bicinchoninic acid assay method. Primary antibodies (CDK2-ab6538, Cyclin A-ab7956, Cyclin E-ab7959, Cyclin D1-ab16663, p27^KIP1-ab7961, Blc-2-ab32124, and cytochrome C-ab133504) were purchased from Abcam (Cambridge, UK).

Western blotting and indirect immunofluorescence were performed using the following primary antibodies: β actin (A5441, Sigma-Aldrich, Ireland); β1 integrin (#1952, Merck Millipore, Ireland); cyclin B1 (#4135), estrogen receptor α (#8644), HER2 (#4290), EGFR (#4267), IGF-1R (#3027), pan AKT (#2920), pan ERK (#4696), pan p-AKT (#4060), pan p-ERK (#4377), p-EGFR (#3777), progesterone receptor A/B (#8757) [all from Cell Signaling Technology Europe, Leiden, Netherlands]; cyclin D1 (ab16663) and N-cadherin (ab19348) [both from Abcam, Cambridge, UK]; E-cadherin (#610181, BD Biosciences, San Jose, CA, USA); GAPDH (sc47724) and vimentin (sc32322) [both from Santa Cruz Biotechnology, Dallas, TX, USA]. Secondary antibodies for western blots were IRDye 800CW or 680RD (LI-COR Biosciences UK Ltd., Cambridge, UK); and the secondary antibody used for β actin indirect immunofluorescence was DyLight 488 donkey anti-mouse (Jackson ImmunoResearch Europe Ltd., Ely, UK).