Pyrimidine Salvage Enzymes Are Essential for De Novo Biosynthesis of Deoxypyrimidine Nucleotides in Trypanosoma brucei
Fig 5
HsDCTD rescues the growth defect in TbTK RNAi and TbTK null cell lines.
A. Growth curves for TK RNAi cells or TK RNAi cells containing a Tet-regulated expression plasmid for HsDCTD. Cell growth was monitored ±Tet for the indicated days. Error bars represent SD for triplicate biological replicates. Inset shows a Western blot of HsDCTD expression ± Tet at 48 h. B. qPCR analysis of TbTK mRNA expression in both the TK RNAi cell line and the TK RNAi HsDCTD rescue line (±Tet 48 h). Error bars represent SEM for triplicate data. Data were normalized to the -Tet control, which is in the background of the single allele TK knockout. C. Growth analysis of TK null cells expressing either FLAG-tagged TbTK (c-null) or FLAG-tagged HsDCTD under the control of the Tet promoter. Error bars represent SD for triplicate biological replicates. Inset shows western blot analysis of the HsDCTD TK null cells 2 days and 5 days after Tet withdraw.