Extended Data Fig. 5: PLCγ1 suppression decreases mitochondrial respiration and enhances cancer cell glycolytic capacity in MEFs.
From: PLCγ1 suppression promotes the adaptation of KRAS-mutant lung adenocarcinomas to hypoxia
a, Immunoblot analysis of the indicated targets in MEF cells with the indicated genotype transfected with either pcDNA3.1-HA-LIC empty vector or pcDNA3.1-HA-LIC-PLCγ1. 24h later, cells were moved in hypoxia (3% O2) and incubated for additional 48h. b, Oxygen consumption rate (OCR) graph of MEF cells of the indicated genotypes. Cells were incubated for 48h in normoxia or hypoxia (3% O2) before seahorse experiment. OCR was determined during sequential treatments (indicated with arrows) with oligomycin, FCCP and rotenone/antimycin (AA) at the indicated time; n = 3 for the normoxia groups and n = 6 for the hypoxia groups. c, Graph showing extracellular acidification rate (ECAR) of MEF cells with the indicated genotype transduced as in (a). ECAR was determined during sequential treatments (indicated with arrows) with glucose (Glc), oligomycin and 2 deoxyglucose (2DG) at the indicated time; n = 6. d, Flow cytometry panels showing mean fluorescent intensity of A549 cells from experiment reported in main Fig. 3i. A549 cells were previously transduced with either an empty vector control (Tet-pLKO-puro, shControl) or 2 different doxycycline-inducible shRNAs against PLCγ1 (shPLCγ1 #1, shPLCγ1 #2), incubated in the presence of doxycycline for 48h, incubated for additional 48h in normoxia or hypoxia (1% O2), stained with LipidTOX and analyzed by flow cytometry. Sh#1: shPLCγ1 #1, Sh#2: shPLCγ1 #2; n = 3. Graphical data are mean ± SD; n, number of biologically independent samples. Statistical source data and unprocessed immunoblots are provided in Source Data Extended Data Fig. 5.
