Journal: The EMBO Journal
Article Title: Limited oxygen in standard cell culture alters metabolism and function of differentiated cells
doi: 10.1038/s44318-024-00084-7
Figure Lengend Snippet: ( A ) Extracellular measurements of 3T3-L1 medium glucose and lactate 24 or 48 h after medium volume change in 12-well plates ( n = 3 technical replicates from n = 3 biological replicates). ( B ) 2-deoxyglucose (DG) uptake after insulin stimulation and 200 µM indinavir (GLUT4 inhibitor) treatment. Cells were cultured in high or low medium for 48 h in 24-well plates prior to the experiment ( n = 6 biological replicates). ( C ) Percentage inhibition of 2-DG uptake after indinavir treatment, calculated from the difference between +/− indinavir treated conditions, as a percentage of -indinavir 2-DG uptake upon 100 nM insulin stimulation. Graph shows the percentage of 2-DG uptake that is GLUT4-dependent (i.e. inhibited by indinavir) ( n = 6 biological replicates). ( D ) Western blot of GLUT1 and GLUT4 in 3T3-L1s after 48 h medium volume change (12-well plate) ( n = 6 biological replicates). ( E ) The pericellular oxygen concentration of primary scAdips cultured with different medium volumes in 96-well plates ( n = 4 biological replicates). AA antimycin A. ( F ) OCR of primary scAdips cultured with different medium volumes in 96-well plates (n = 4 biological replicates). Data information: Data were represented as mean ± SEM ( A – D ). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 by two-way ANOVA with Šidák correction for multiple comparisons ( A . B ), or by paired two-tailed Student’s t -test ( C ).
Article Snippet: Indinavir , Sigma-Aldrich , Cat # SML0189.
Techniques: Cell Culture, Inhibition, Western Blot, Concentration Assay, Two Tailed Test