FIG. 7.

Mitochondria were prepared from MTF (A) and HΔC (B) cells cultured without doxycycline. The mitochondria were solubilized with 0.5% NP-40 (total) and separated into pellet (P1) and supernatant (S1). The P1 fractions were treated with nuclease S7 or DNase-free RNase A and then separated again into pellet (P2) and supernatant (S2). Each sample was analyzed by Western blotting with anti-human TFAM, anti-P32, anti-mtSSB, and anti-HA antibodies. mtDNA in total, P1, and S1 were detected by PCR. Human TFAM (arrowhead), mouse TFAM-HA (double arrowheads), and human TFAM-ΔC-HA (triple arrowheads) are indicated.