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Fig. 4 | BMC Biology

Fig. 4

From: The mitochondrial carrier pathway transports non-canonical substrates with an odd number of transmembrane segments

Fig. 4

Mpc2 and Mpc3 import depends on small TIM chaperones. a Radiolabeled Mpc2 and Mpc3 were imported at 25 °C into wild-type (WT) mitochondria, mitochondria with the mutant form Tim10-L26Q, mitochondria lacking Tim8 and Tim13, or mitochondria affected in Tim10, Tim8, and Tim13 [26]. Samples were analyzed by BN-PAGE and autoradiography. Quantification of import and assembly efficiency; the efficiency into Tim10-WT/tim8Δtim13Δ mitochondria after 10 min was set to 100% (control); n = 3 for Mpc2 import, n = 4 for Mpc3 import; error bars: SEM. b AAC (upper panel) and F1β (lower panel) were imported at 30 °C into wild-type or tim12-21 mutant mitochondria, followed by BN-PAGE (AAC) or SDS-PAGE (F1β) analysis and autoradiography. m, mature form. c Mpc2 (upper panel) and Mpc3 (lower panel) were imported at 30 °C into wild-type or tim12-21 mutant mitochondria and analyzed by BN-PAGE and autoradiography. d Mpc2 or Mpc3 were imported at 30 °C into wild-type or tim12-21 mutant mitochondria. Mitoplasts were generated by hypo-osmotic swelling, treated with proteinase K, and analyzed by SDS-PAGE and autoradiography (upper panel) or Coomassie Blue R-250 staining (Coom.) as a loading control (lower panel). Quantification (right panel) of membrane potential (Δψ)-dependent import yield after 10 min relative to the WT control; n = 3; error bars: SEM. In all experiments, non-imported precursors were degraded with proteinase K

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