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For purposes of normalization the blots were also stained with a monoclonal anti-actin antibody in a dilution of 1 400 (Amersham).
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Two lanes in each blot were also stained for α1 subunits; all images were taken after 10 min. Experiments were performed three times, and each subunit was detected on one blot in duplicates.
Sections were also stained by immunofluorescence.
The nucleus was also stained.
The blot was also re-probed and corrected by staining with β-actin antibody (Sigma, St Louis, MO, USA).
Some samples were also Gram stained.
Blots were stained with methylene blue staining to confirm efficient transfer and equal loading.
Blots were then stained with Ponceau S and photocopied.
For detection of PHLDA1 expression, blots were stained with 1 1000 dilution of anti-PHLDA1 antibody (EPR6674, abcam®).
For detection of PDK4 expression, blots were stained with 2 µg/mL dilution of anti-PDK4 (Novus07049, Novus Biologicals).
All blots were stripped and stained with HRP conjugated anti β-actin antibody (Sigma, 1∶1000).
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