Lamina c

Total protein was extracted using RIPA lysis buffer from Yeasen (China). Cytoplasmic and nuclear protein extraction was carried out using the Nuclear and Cytoplasmic Protein Extraction Kit (Yeasen, China). ACTIN and LaminA/C were used as internal controls for cytoplasmic and nuclear proteins, respectively. Primary antibodies used in this study were, c-Myc(ab32072, Abcam, USA), PCNA (A0264, ABclonal, China), SRSF1 (sc-33652, Santa Cruz Biotechnology, China), β-catenin (51067-2-AP, Proteintech, USA), cyclin D1(ab134175, Abcam, USA), LaminA/C (A0249, ABclonal, China), and ACTIN (AC026, ABclonal, China).

Cells were lysed with RIPA peptide lysis buffer (P0013B; Beyotime, Shanghai, China) and centrifuged at 13,000 g for 10 min. The resulting supernatants were used for western blotting. Nuclear and cytoplasmic proteins were extracted using the Nuclear and Cytoplasmic Protein Extraction Kit (P0028; Beyotime, Shanghai, China) according to the manufacturer’s instructions. Protein concentrations were determined using an Enhanced BCA Protein Assay Kit (P0010S; Beyotime). Protein extracts were separated by 10% SDS-PAGE and probed with the following antibodies (1:1000): E-cadherin (cat. no. 3195; Cell Signaling Technology, Danvers, MA, USA), N-cadherin (cat. no. 13116; Cell Signaling Technology), vimentin (cat. no. 5741; Cell Signaling Technology), Snail (cat. no. 3879; Cell Signaling Technology), Twist (cat. no. A7314; ABclonal, Wuhan, China), ZEB1(cat. no. ab203829; Abcam), Slug (cat. no. 9585; Cell Signaling Technology), β-catenin (cat. no. 8480; Cell Signaling Technology), β-actin (cat. no. AC026; ABclonal), LaminA/C (cat. no. 4777; Cell Signaling Technology). Antibody binding was revealed using an HRP-labelled goat anti-rabbit IgG (H + L) (A0208; Beyotime). The signals were visualised using a GelDoc XR+ imaging system (Bio-Rad). Integrated density of E-cadhein and N-cadherin was analyzed by ImageJ software to quantified E-cadhein/ N-cadherin ratio.