Histone h3 ab1791

Mitochondria were prepared as previously described (Shephard et al., 2014 (link)). The quality of the crude fractions was confirmed using standard Western blotting techniques with nuclear, mitochondrial and cytoplasmic markers (Histone H3, ab 1791 (Abcam); COX IV ab16056 (Abcam); and HSP-90 ab13495 (Abcam) respectively). Sub-fractions were confirmed using outer membrane, inner membrane and inter-membrane space markers.

The E-cadherin–GFP expression construct was a gift from Alpha Yap^{50 (link)} (University of Queensland, Queensland, Australia). Wild-type E-cadherin was generated by PCR amplification of the corresponding cDNA fragments using E-cadherin–GFP as a template. The E-cadherin tyrosine-to-alanine (⁷⁵³YYY^755→753AAA⁷⁵⁵) mutant was generated by site-directed mutagenesis using wild-type E-cadherin as a template. The correct clone sequence was verified by sequencing. Lentiviral-TOP-dGFP-reporter plasmids were obtained from Addgene (Addgene plasmid 14715, Cambridge, MA, USA). A pLKO.1-shRNA encoding an shRNA with a scrambled sequence or sequences targeting human Sox15 and E-cadherin, purchased from the National RNAi Core Facility, Taiwan, was introduced into HEK293T cells using the lentiviral packaging vectors pMD.G and pCMV_8.91. Antibodies against the following proteins were used: E-cadherin (610181; BD Biosciences, San Jose, CA, USA); histone H3 (Ab1791) and ABCG2 (Ab3380) (all from Abcam, Cambridge, MA, USA); active β-catenin (ALX-804-260/1; dephosphorylated at Ser33/37; Enzo Life Sciences, Farmingdale, NY, USA); total β-catenin (C2206), β-actin (A5441) and TCF4 (T5817) (all from Sigma-Aldrich, St Louis, MO, USA); CD133 (3663) and phosphorylated β-catenin (phosphorylated at Ser33/37/Thr41) (9561) (all from Cell Signaling Technology, Beverly, MA, USA).