Total protein in cultured cells was extracted by RIPA Lysis Buffer (Beyotime), and protein concentration was measured using BCA Protein Assay Kit (Beyotime). Proteins were denatured in loading buffer by boiling at 100 °C for 10 min, and denatured protein samples were subjected to Western blotting assay [20 (link)]. The antibodies used were presented in Table 3 . ECL™ Western blotting system (Merck, Darmstadt, Germany) was used to detect protein signals and densitometric quantification was performed using ImageJ software (NIH, Bethesda, MD, USA).
The antibodies used in this study
| Name | Cat. no. | Dilution ratio | Source |
|---|---|---|---|
| LDHA | AF0216 | 1:1000 | Beyotime |
| PDK1 | AF7707 | 1:1000 | Beyotime |
| β-actin | AA128 | 1:1000 | Beyotime |
| HRP-labelled anti-Rabbit IgG | A0208 | 1:1000 | Beyotime |
| HRP-labelled anti-Mouse IgG | A0216 | 1:1000 | Beyotime |
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