Fig. 2
NIS signal in A549-LN cells is dose-dependently decreased following oxidative phosphorylation inhibition. a Radioactivity uptake in A549-LN cells in vitro at 60 min from the addition of 99mTc at 50 kBq/mL and vehicle or IACS-10759, at specified concentrations. Decay-corrected radioactivity counts were normalised to protein concentrations assayed in samples of lysates used in radioactivity measurements. For presentation, individual data points were normalised to the mean of control. b Summarised results of trypan blue exclusion assay performed in separate set of cell cultures at 60 min from the beginning of drug conditioning as described in a. c LC-MS measurements of selected intra- and extracellular metabolites extracted at 60 min from the beginning of IACS-10759 conditioning. Volume of the extraction buffer was adjusted to 1 mL per 2.5 × 106 cells according to cell counts obtained from replicate plates, conditioned as per plates used in the extraction. d LC-MS measurements of adenine nucleotide concentrations collected in experiments described in c. e Energy charge ratio (ECR) calculated for LC-MS experiments described in c and d from the formula ECR = ([ATP] + 0.5 [ADP])/([ATP] + [ADP] + [AMP]), where [metabolite] is the metabolite’s peak area. Ordinary, one-way, multiple comparisons ANOVA was performed to test statistical significance of the results, and P values calculated using Dunnett’s multiple comparisons test, evaluating all conditions versus the control at each respective time point were presented. P value classifications are summarized as follows: *, P ∈ (0.01–0.05 〉; **, P ∈ (0.001–0.01 〉; ***, P < 0.001. Only statistically significant (P < 0.05) results are presented. All experiments used n = 3 or n = 6 (for trypan blue assay) biological replicates, each represented by a data point. Error bars represent one standard deviation