Fig. 2
From: Genetic dissection of the mitochondrial lipoylation pathway in yeast
Analysis of lipoylation deficient strains expressing Fam1-1 (mitochondrially targeted Fam1/Faa2). All plasmids transformed into the different strains are based on the YEp352 yeast episomal plasmid backbone. a Growth assays of wild-type +YEp352, Δhtd2 + YEp352, Δhtd2 + YEp352mtFam1-1, Δhtd2Δlip2 + YEp352, Δhtd2Δlip2 + YEp352mtFam1-1, Δhtd2Δgcv3 + YEp352, and Δhtd2Δgcv3 + YEp352mtFam1-1. 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 a SCD-URA plate as a general growth control and on SCG-URA media supplemented with LA or C8. Plates were incubated at 30 °C for 4 days. b Western blot analysis of extracts of mtFam1-1 complemented strains. Whole cell extracts were collected from cells harboring YEp352mtFam1-1 or YEp352 as a negative control, after 16 h of growth on YPG media at 30 °C. Analysis was done by probing with anti-LA serum and anti-actin serum as a loading control. Whole cell extracts from wild-type+YEp352, Δhtd2 + YEp352, Δhtd2 + YEp352mtFam1-1, Δhtd2Δlip2 + YEp352, and Δhtd2Δlip2 + YEp352mtFam1-1 grown without supplements (−) or with 100 μM C8 supplementation were analyzed with anti-LA serum. See Additional File Fig. A1 for further data