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Primary Ab dilution-1:1000 dilution |
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Primary Ab dilution-1:1000 dilution |
Publication protocol
Cells and tumour tissue samples were prepared and subjected to western blotting as previously described.29 All primary antibodies were used at 1/1000 apart from anti-β-actin, which was diluted at 1:10,000. Anti-horseradish peroxidase-conjugated antimouse, anti-rabbit and anti-rat IgG secondary antibodies were used at a concentration of 1:2000. Densitometry data from western blot experiments were generated using the ImageJ software (NIH, USA) by measuring grey intensities of lanes corresponding to the protein of interest relative to the intensity of the lane corresponding to the loading control. Data are expressed as a fold change compared to control. It is indeed possible that some densitometry data (fold increase) were overestimated in cases where control values were nearly undetectable. This method is semi-quantitative and absolute data should indeed be interpreted with caution. Where available (e.g. measurements of p62) quantitative ELISA was used.
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The HDAC6 inhibitor C1A modulates autophagy substrates in diverse cancer cells and induces cell death.
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