Fig. 8
From: Genetic dissection of the mitochondrial lipoylation pathway in yeast
Complementation of Δlip2and Δlip3 yeast strains by human LIPT1 and LIPT2. All plasmids transformed into the different strains are based on the YEp352 yeast episomal plasmid backbone. a Growth assays: wild-type+YEp352, Δlip3 + YEp352, Δlip3 + YEp352hsLIPT1, Δlip2 + YEp352, and Δlip2 + YEp352hsLIPT2. Cells were grown for 16 h, adjusted to an OD600 of 0.5 and spotted as 1× 1/10×, 1/100×, and 1/1000× serial dilutions on SCD-URA media as a general growth control and on SCG-URA media. Plates were incubated at 30 °C for 4 days. b Western blot analysis of cell extracts of wild-type+YEp352, Δlip3 + YEp352, Δlip3 + YEp352HsLIPT1, Δlip2 + YEp352, and Δlip2 + YEp352HsLIPT2 with anti-LA serum. c Growth assays of wild-type, Δgcv3Δlip3 + YEp352-LplAK133L, Δgcv3Δlip3 + YEp352-LplA, Δgcv3Δlip3 + YEp352-LIPT1 + YCp22-LplAK133L, Δgcv3Δlip3 + YEp352-LIPT1 + YCp22-LplA. Cells were spotted on a SCD-URA-TRP media as a general growth control and on SCG-URA-TRP media without supplements or supplemented with 100 μM LA or 100 μM C8. Plates were incubated at 30 °C for 4 days