Anti β actin sc 47778hrp

The target gene fragments were amplified by PCR and were inserted into pcDNA3.0 (Invitrogen) to construct eukaryotic expression vectors. The target sequence of BEND5 shRNA was 5'-GCAAATACGTCGTCCTATT-3'. The human breast cancer cell lines MDA-MB-231 and ZR75-1, and human embryonic kidney cell line HEK293T were purchased from the American Type Culture Collection. Anti-BEND5 (20931-1-AP), anti-HEY2 (10597-1-AP), anti-E-cadherin (20874-1-AP), anti-N-cadherin (22018-1-AP), anti-Vimentin (10366-1-AP), and anti-MAML1 (55493-1-AP) were purchased from Proteintech. Anti-HES1 (ab71559) was purchased from Abcam. Anti-BEND5 (NBP2-26211) was from Novus Biologicals. Anti-RBPJ (sc-271128), anti-Notch1 (sc-373891), and anti-β-actin (sc-47778HRP) were purchased from Santa Cruz Biotechnology. Anti-FLAG (A8592), anti-FLAG M2 agarose (A2220), anti-c-MYC gel (E6654), and anti-c-MYC-peroxidase (A5598) were from Sigma-Aldrich. Dulbecco's modified Eagle's medium (DMEM, 12491-015) was purchased from Gibco. VigoFect reagent (T001) was purchased from Vigorous Biotechnology. CCK8 reagent (CK04) was purchased from Dojindo. Paraformaldehyde (BL539A) was purchased from Biosharp.

Human breast cancer cell lines MCF7 and ZR75-1 were obtained from the American Type Culture Collection (ATCC, USA). Cells were cultured in DMEM (Dulbecco's modified Eagle's medium, Invitrogen, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (Gibco, Carlsbad, CA, USA) and 1% penicillin-streptomycin solution 100X (Corning, CA, USA) in a humidified atmosphere with 5% CO₂ at 37 °C. Specific antibodies against PGM5 (PA5-48880) was obtained from Invitrogen. Anti-p21 (ab227443), anti-p53 (ab131442), anti-BAX (ab216494), anti-FAK (ab40794), and anti-LDHB (ab112996) were purchased from Abcam. Antibodies against cyclin B (#4138), cyclin D1 (#2922), Bcl-2 (#2876), phos-FAK (Y397) (#8556), MMP-2 (#4022), MMP-9 (#3852), and LDHA (#3582) were obtained from Cell Signaling. Anti-E-cadherin (sc-8426), anti-vimentin (sc-6260), anti-N-cadherin (sc-8424), and anti-β-actin (sc-47778HRP) antibodies were purchased from Santa Cruz Biotechnology.

The eukaryotic expression constructs for GATA1 were generated by cloning PCR-amplified full length sequences into pcDNA3 (Invitrogen). GST-fusion protein encoding vectors were constructed by inserting PCR-amplified sequences into pGEX-KG (Amersham Pharmacia Biotech). The Bcl-XL promoter luciferase reporters were obtained by cloning PCR-amplified promoter fragments into pGL4-basic vector (Promega). The mutated Bcl-XL promoter luciferase reporters were constructed by recombinant PCR. Lentiviral vector of GATA1 was constructed by cloning PCR-amplified full length sequences into pCDH (System Biosciences). The short hairpin RNAs (shRNAs) targeting GATA1 and Bcl-XL cDNA were inserted into PSIH-H1-puro (System Biosciences). The small interfering RNAs sharing the same targets with shRNAs were synthetized from GenePharma (Shanghai, China). The sequences for siRNAs and shRNAs are listed in Table S4a.
Specific antibodies against GATA1 (ab28839) and Bax (ab32503) were purchased from Abcam. Anti-Bcl-XL (#2764), anti-cleaved PARP (#5625), anti-cleaved caspase 3 (#9664), and anti-cleaved caspase 9 (#9505) were from Cell Signaling Technology. Anti-β-actin (sc-47778HRP) was from Santa Cruz Biotechnology. Gemcitabine (T0251) was obtained from Targetmol.