Fig. 5: AnkJ rescues Ceg14-induced ATP depletion in mammalian cells.

a AnkJ rescues Ceg14-induced cell growth arrest. HEK293T cells of 50% confluence were transfected to express the indicated proteins. Images were acquired 24 h post-transfection (left panels). The expression of the relevant proteins was probed with antibody specific for Flag or HA. GAPDH was probed as a loading control (right panels). V, empty vector. Data shown were a representative of three independent experiments with similar results. Scale bar, 20 μm. b AnkJ counteracted cell viability reduction induced by Ceg14. Samples expressed combinations of proteins were used to determined cell viability. c AnkJ rescued ATP depletion induced by Ceg14. HEK293T cells were transfected to express the indicated proteins for 24 h, and lysates of equivalent number of cells were used to measure ATP. The level of ATP in cells transfected with the empty vector was used as the baseline for comparison. Data shown (mean ± s.e.) in (b, c) were from three replicates. Unpaired two-tailed Student’s t-tests were performed. d, e Modulation of cellular ATP levels in Raw264.7 (d) and D. discoideum (e), by Ceg14 and AnkJ during L. pneumophila infection. ATP levels were determined in cells infected with the indicated bacterial strains at the indicated time points. The ATP level of uninfected cells at the 5 min time point was used as a control. Results (mean ± s.e.) shown were from three independent experiments. Ordinary one-way ANOVA with multiple comparisons was performed for data analysis. Source data are provided as a Source Data file.