For immunoblot analysis, cells were lysed in RIPA lysis buffer (Thermo Fisher Scientific, #89901) supplemented with protease inhibitor cocktail (#87786), and total cell lysates were collected for further uses. In some experiments, nuclear and (or) cytoplasmic proteins were extracted using the NE-PER Nuclear and Cytoplasmic Extraction Reagents (Thermo Fisher Scientific, #78833) according to the manufacturer’s instructions. The cell lysates were subjected to immunoblot assay using primary antibodies against Snail (#3895; 1:1000), Slug (#9585; 1:1000), Cyt-c (#4280; 1:1000), caspase 3 (#9665; 1:1000), caspase 9 (#9508; 1:1000), cleaved caspase 3 (#9661; 1:500), cleaved caspase 9 (#52873; 1:500), p53 (#2524; 1:1,000), Bax (#2772; 1:1000), Puma (#24633; 1:1000), pan-acetyl-Lys (pan-AcK; #9441; 1:500), CBP (#7389; 1:1000), p300 (#70088; 1:1000), ubiquitin (#3936; 1:1000), HDAC1 (#5356; 1:1000), vimentin (#5741; 1:1000), histone H3 (#4499; 1:2000), HA-tag (#3724S; 1:2000), β-tubulin (#2128; 1:2000) (all from Cell Signaling Technology), E-cadherin (BD Biosciences, #610181; 1:5000), p21 (#ab7903; 1: 200), MDM2 (#ab16895; 1:500), phospho-Ser/Thr (#ab17464; 1:1000) (all from Abcam), FLAG (#F3165; 1:1000), importin β (Thermo Fisher Scientific, #MA3-070), and β-actin (#A5316; 1:10,000) (both from Sigma-Aldrich), followed by incubation with appropriate horseradish peroxidase (HRP)–conjugated secondary antibodies. Blots were detected by enhanced chemiluminescence (Thermo Fisher Scientific, #32106). For IP assay, cells were lysed in IP lysis buffer [50 mM tris-HCl, 150 mM NaCl, 1 mM EDTA, and 1% NP-40 (pH 7.4)] containing protease inhibitor cocktail for 20 min on ice. The cell lysates were sonicated, clarified, and incubated with antibodies against control immunoglobulin G, FLAG (1:100), Snail (1:100), HDAC1 (1:100), or p53 (1:100), followed by incubation with precleared Protein A/G agarose beads (Santa Cruz Biotechnology, #sc-2003). The immunocomplexes were subjected to immunoblot analysis using antibodies against ubiquitin, HA, pan-AcK, phospho-Ser/Thr, CBP, p300, FLAG, or p53. For His pulldown assay, GST-CBP-HAT, His-Snail-WT, and His-Snail-R174A mutant recombinant proteins were expressed and purified from E. coli (BL21). The bead-bound His-tagged proteins were preincubated with various concentrations of CYD19 for 15 min at 4°C on a rotator, and eluted GST-CBP-HAT protein was added to the reaction mixtures and incubated for another 2 hours. The beads were collected, extensively washed, eluted, electrophoresed, and subjected to Coomassie staining. In some experiments, His-Snail-WT and His-Snail-R174A mutant recombinant proteins were immobilized to Ni-NTA agarose and incubated with whole lysates of HEK293T cells for 3 hours (34). After extensive washes, the bound proteins were eluted with SDS sample buffer, resolved by SDS–polyacrylamide gel electrophoresis, and analyzed by immunoblotting.

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