Journal: JCI Insight
Article Title: Mitochondrial reprogramming by activating OXPHOS via glutamine metabolism in African American patients with bladder cancer
doi: 10.1172/jci.insight.172336
Figure Lengend Snippet: ( A ) Protein expression of OXPHOS cocktail — mitochondrial complex V (ATP5A), III (UQCRC2), II (SDHB), IV (MT-CO2), and I (NDUFB8) — in EA BLCA (UM-UC-3, T24) and AA BLCA (UM-UC-1, SCaBER) cell lines. ( B ) Mitochondrial complex activity in AA BLCA — UM-UC-1 ( n = 3), SCaBER ( n = 3) — cell lines compared with EA BLCA — UM-UC-3 ( n = 3), UM-UC-5 ( n = 3) — cell lines; optical density (OD) of complex I activity was measured using specific substrates and normalized with the citrate synthase activity measured by colorimetric assay ( ****P < 0.0001). ( C ) Box-and-whisker plot represents ATP production in AA BLCA — UM-UC-1 ( n = 7), SCaBER ( n = 7) — and EA BLCA — UM-UC-3 ( n = 7), T24 ( n = 7) — cell lines measured by Seahorse assay. Data were normalized using cell number measured by CellTiter-Glo (*** *P < 0.0001). ( D and E ) Scatter plot represents the tumor growth (orthotopic) of AA BLCA (UM-UC-1) and EA BLCA (UM-UC-3) upon IACS-010759 (5 mg/kg) treatment compared with vehicle control. Luciferase measurements were performed over a period of 21 days — UM-UC-1: days 7 and 14 for Control ( n = 7), IACS-010759 treatment ( n = 8); day 21 for Control ( n = 6), IACS-010759 treatment ( n = 8); and UM-UC-3: day 7 for Control ( n = 8), IACS-010759 treatment ( n = 8); day 14 for Control ( n = 8), IACS-010759 treatment ( n = 7); day 21 for Control ( n = 7), IACS-010759 treatment ( n = 7) (unpaired t test for each time point; ***P < 0.001). ( F ) Plot represents the tumor growth (s.c.) from AA BLCA (SCaBER) cell line–derived xenografts treated with IACS-010759 (5 mg/kg; n = 5) compared with vehicle control ( n = 5) (unpaired t test for each time point; **P < 0.01 ; *P < 0.05 ) . ( G ) Protein expression of NDUFB8 (mitochondrial complex I protein) in EA ( n = 4; E1 to E4) and AA BLCA ( n = 4; A1 to A4) tissues measured by Western blot analysis. Significance was determined by unpaired 2-tailed student t test.
Article Snippet: UM-UC-3, T24, J82, and SCaBER (HTB-3) cell lines were purchased from ATCC, and UM-UC-1 and UM-UC-5 cell lines were purchased from Sigma-Aldrich.
Techniques: Expressing, Activity Assay, Colorimetric Assay, Whisker Assay, Control, Luciferase, Derivative Assay, Western Blot