Fig. 4: Mitochondrial ATP-linked respiration was followed by TM4SF5-mediated ALB uptake.
Huh7-KO3#309 cells were transduced using a lentivirus for control empty vector, TM4SF5WT or TM4SF5A132V expression. On the basis of OCR or ECAR analyses, ATP-linked and maximal respiration as parameters of mitochondrial function and glycolysis or glycolytic capacity for glycolytic functions were calculated and plotted at mean ± s.e.m. values. Each data point in the graphs from the OCR and ECAR measurements represent a mean value of a triplicate experiment. a–d Cells were then replenished with ALB to GLU-free SFM (a) or to GLU-containing SFM (b) without or with EIPA (10 μM, a, b and d) or CQ (25 μM, c) treatment for 15 h before a mitochondria stress test to measure their OCR (a–c) or before measurement of ECAR (d). e, f Cells were replenished with single amino acids to GLU-free (e) or GLU-containing (f) SFM with or without EIPA treatment for 15 h, before OCR analysis. g Cells were replenished with Phe at different concentrations to GLU + SFM for 15 h before OCR analysis. h Huh7KO-TM4SF5WT cells were replenished with or without ALB alone or each amino acid to GLU + SFM for 15 h. Relative FITC-ALB uptake was measured in replated on collagen I as an index of macropinocytosis. P values were calculated via one-way ANOVA or unpaired Student’s t-tests, and P < 0.05 was considered statistically significant. *, **, *** and **** depict P < 0.05, 0.01, 0.001 and 0.0001, respectively. Data represent three independent experiments. See also Supplementary Fig. 5.
