FIGURE 2.

Effect of Lig3 knockdown on mtDNA copy number. Six shRNA constructs (2842-1 through 6) were generated, and their effects on Lig3 expression and mtDNA copy number were studied. A, Western blots of whole cell lysates from HeLa cells infected with shRNA constructs 2842-1 through 6. β-Actin, loading control. B, Western blotting of serial dilutions of the mitochondrial lysate of the WT HeLa cells and undiluted mitochondrial lysate of HeLa/2842-2. Mitochondrial HSP60, loading control. C, QSBN of the total DNA from HeLa cells and cells infected with knockdown constructs. Total DNA was digested with BamHI and processed as described under “Experimental Procedures.” Mito, mitochondria. D, quantitation of the QSBN in C. E, quantitation of mtDNA copy number in WT HeLa cells and HeLa/2842-2 by TaqMan qPCR. Error bars, S.E., n = 3.