Anti cyclin d1

Primary antibodies: anti-YB-1 raised against the region 1 to 100 of YB-1 protein (12148 Abcam, Cambridge, UK); anti-PABP1 (poly(A)-binding-proteiin 1) (clone 10E10, Sigma-Aldrich); anti-Myc (SAB21084786, Sigma-Aldrich, Saint Louis, MI, USA); anti-cyclin D1 (GT8912, Genetex, Irvine, CA, USA); anti-cyclin A2 (GT2547, Genetex, Irvine, CA, USA); anti-GAPDH (6C5 Santa Cruz Biotechnology, Dallas, TX, USA); anti-PARP1 (Cell Signalling, Danvers, MA, USA); anti-RNH1 (436–450, Sigma-Aldrich, Saint Louis, MO, USA) anti-β-tubulin (H-235, Santa Cruz Biotechnology, Dallas, TX, USA); anti-actinin (AT6/172, Abcam, Cambridge, UK); anti-p21 WAF (1D21 Cell Signaling, Danvers, MA, USA); anti-∆Np63α (4A4) (sc-8431 Santa Cruz Biotechnology, Dallas, TX, USA).
Secondary fluorescent antibodies: Alexa Fluor 488 anti-rabbit (Thermo-Fisher Scientific, Waltham, MA, USA); Cy3 anti-mouse (Jackson ImmunoResearch, Cambridge, UK); DAPI (Sigma-Aldrich, Saint Louis, MO, USA).

After the addition of 100 nM of calcitriol (Cayman Chemical, Ann Arbor, Michigan) for 48 h or transfection for 24 h (overexpression) or 72 h (RNAi), HT29 or SW480 cells were washed with PBS briefly, and the preparation of total cell lysates was as stated before [20 (link)]. Cellular proteins of the whole cell lysates were separated and resolved by SDS–PAGE and electrophoretically transferred onto polyvinylidene fluoride membranes (Perkin Elmer Life Sciences, Inc., Waltham, MA). Western blotting was performed as previously described [20 (link)] with the following primary and secondary antibodies: MED28, β-catenin, and p-GSK3β (Ser9) antibodies from MilliporeSigma (Burlington, MA); fibronectin, E-cadherin, MMP2, and MMP9 antibodies from Abcam (Waltham, MA); anti-Flag-tag and anti-p21 from Proteintech Group, Inc. (Rosemont, IL); antibodies for vimentin, Snail, Slug, Twist, ZEB2, proliferating cell nuclear antigen (PCNA), α-tubulin, β-actin, vinculin, and GAPDH from Santa Cruz Biotechnology (Santa Cruz, CA); and anti-c-Myc, anticyclin D1, and goat antirabbit or antimouse-conjugated horseradish peroxidase secondary antibodies from GeneTex, Inc. (Irvine, CA).