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. 2021 Jun 30;13(13):3285. doi: 10.3390/cancers13133285

Figure 5.

Figure 5

Effect of CerK silencing on atrogin-1/MAFbx and CerK protein and mRNA expression in C2C12 myotubes. (A–D): cell lysates for CerK and atrogin-1/MAFbx protein expression (A) and total RNA for atrogin-1/MAFbx (C) or CerK (D) mRNA expression were obtained from C2C12 myotubes transfected with scrambled shRNA (SCR-shRNA) or shRNA specific to CerK (CerK-shRNA), as described in Methods. (A,B): lysates (40 µg) from C2C12 myotubes transfected with SCR-shRNA or CerK-shRNA were subjected to SDS-PAGE and immunoblotted with anti-CerK or anti-atrogin-1/MAFbx antibodies. A blot representative of the densitometric analysis is shown. Normalized data (means ± SEM), to β-actin band, are reported (Student’s t-test, * p < 0.05 vs. SCR-shRNA). (C,D): total RNA was purified, reverse transcripted and analyzed as reported in Methods using specific primers for atrogin-1/MAFbx (C) or CerK (D). Data are presented as fold change (mean ± SEM) of at least 3 independent experiments (Student’s t test,* p < 0.05, ** p < 0.01 vs. SCR-shRNA).