Exploring the Role and Mechanism of pAMPKα-Mediated Dysregulation of Brf1 and RNA Pol III Genes

Figure 6

The role of Brf1 alteration in transcription of Pol III genes. (a) Immunoblotting analysis of Brf1 and pAMPKα protein levels. A549 cells were treated with AMPK inhibitor, S7306 (12 h with 10 μM), and MNNG (1 h with 4 μM). (b) RT-qPCR analysis of Brf1 mRNA levels in A549 cells treated with S7306 (12 h with 10 μM) and MNNG (1 h with 4 μM). (c) Immunoblotting analysis of Brf1, pAMPKα, and AMPKα protein levels in MNNG-treated A549 cells after siRNA-mediated knockdown of Brf1, compared to mm siRNA as control (siCon). (d) RT-qPCR analysis of Brf1 mRNA levels in A549 cells which were transfected with mmRNA or Brf1 siRNA to knock down Brf1. (e, f) RT-qPCR analysis. Pol III gene transcription in A549 cells was transfected with Brf1 siRNA or mmRNA for 48 h and then treated with MNNG (4 μM) for 1 h. All error bars represent the SD of at least three independent experiments. values were determined by a two-tailed -test. , .

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