Fig. 1

NIS signal in HEK293T-SN cells is reduced in response to drugs targeting NaK ATPase, glycolysis and oxidative phosphorylation. a Radioactivity uptake in HEK293T-SN cells in vitro at 30, 60, and 120 min, respectively, after the addition of [¹⁸F]TFB at 50 kBq/mL and drugs at specified concentrations. Decay-corrected radioactivity counts were normalised to cell numbers estimated using replicate plates, conditioned for 120 min as the cell cultures used in radioactivity uptake assay, but without added radioactivity. For presentation, individual data points were normalised to the mean of control. b Results of trypan blue exclusion assay performed at 120 min from the beginning of conditioning as specified in a. c LC-MS measurements of selected intra- and extracellular metabolites at 120 min from the beginning of drug conditioning. Volume of extraction buffer was adjusted to 1 mL per 2.5 × 10⁶ cells according to cell counts obtained from replicate plates, conditioned for 120 min as per plates used in the extraction. d LC-MS measurements of cell adenine nucleotides 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 biological replicates, each represented by a data point. Error bars represent one standard deviation.