Heart and cell homogenates (30 ?g protein), or isolated cytosolic (20 ?g protein), mitochondrial (15 ?g protein), and nuclear (20 ?g protein) fractions were electrophoresed on a 4–15% Mini-Protein TGX gel using a Mini-PROTEAN electrophoresis chamber (Bio-Rad), and proteins were transferred to a PVDF membrane using a Criterion Trans-blot System (Bio-Rad). Membranes were blocked with 5% non-fat dry milk (NFDM) in 50 mM Tris-HCl (pH 7.5) / 150 mM NaCl (TBS), washed with TBS-0.1% Tween 20 (TBST) and TBS, and incubated overnight at 4°C with antibody clones listed in Table C of S1 Table. Antibodies from Santa Cruz were used at 1:200 dilutions, and all other antibodies were used at 1:1000 dilution in TBST-5% NFDM. Membranes were washed with TBST and TBS, incubated with HRP-conjugated secondary antibody (1: 10,000 dilution, Southern Biotech, Birmingham AL), and images were acquired by using an Image Quant LAS4000 system (GE Healthcare, Pittsburgh MA). Immunoblots were subjected to Ponceau S staining to confirm equal loading and transferring of samples. Densitometry analysis of protein bands was performed using a Fluorchem HD2 Imaging System (Alpha Innotech, San Jose CA), and normalized against GAPDH (tissue homogenates and cytosolic fractions), COIV (mitochondrial fractions) or Lamin B (nuclear fractions).
The number of cancer cases diagnosed around the world each year has risen by almost a third in the past decade as increasing life expectancy means millions of more people will be diagnosed in their lifetime.
Researchers said there was also cause for concern in the rise in skin, lung and colorectal cancers, which could largely be prevented by addressing lifestyle habits like smoking and diet.
The Global Burden of Disease Cancer Collaboration audits diagnosis rates and deaths of 29 types of cancer around the world each year.
Its latest report, published in the JAMA Oncology journal, shows there were 17.2 million cancer cases, and 8.9 million cancer deaths around the world in 2016.
This is an increase of 28 per cent in new cancers between 2006 and 2016, much of which is attributed to an expanding and ageing population as well as screening and diagnostics which can catch cancers sooner.
However, the biggest growth has been seen in low and middle income countries – emerging markets which have been targeted by tobacco giants and an influx of cheap convenience foods in recent decades.
“This highlights the importance of focusing tobacco control efforts on lower socio-demographic index countries (SDI),” the authors wrote. “To avoid these countries’ having to experience the same tragedy of unnecessary tobacco related deaths that many high SDI countries have had to face.”.