Smad4 antibody

ChIP was executed with the ChIP assay kit (Beyotime) using the Smad 4 antibody, Smad 4, and anti-IgG antibodies (Cell Signaling Technology). The ChIP DNA was extracted with the DNA Purification Kit (Beyotime) and the samples were then subjected to qPCR amplification with primers spanning the protein-binding sites. The primers used were for miR-26a (F: 5′-GAGGCCTGATGGAGCCGTGGGGACC-3′, R: 5′-AGAG CCACAGCAGGCGGAAAGCCAGATGCCACAGG-3′), miR-375 (F: 5′-TTTCCTAGAACTGCTGTCTTGTCCCATCGCC CACA-3′, R: 5′-AAGAACCCACCCTTTCCTCATCAGAACCAC TTTGC-3′), and Ngn3 (F: 5′-TTAGGCTTTGGATTCTTCATAG TTCTGATAGGATTTGC-3′, R: 5′-AAGTGTCTTCTGGTCCC AGGAATGGAGGCGTAAGG-3′).