Extended Data Fig. 1: Referred to Fig. 1.
From: Primary cilia sense glutamine availability and respond via asparagine synthetase
a, Left: western blot for IFT88 of total cell lysates from MEFIft88 and MEFCtrl generated by CRISPR/Cas9 technology. Right: Representative IF images of MEFIft88 and MEFCtrl. Cilia (ARL13B, green), nuclei (DAPI, blue). Scale bar: 10 µm. Arrows indicate cells in the insets. b, Growth curves in MEFIft88 and MEFCtrl cultured in DMEM + 10% FBS for 8, 24, 48 hours. c, Analysis of OCR measurement in one representative experiment in MEFIft88 and MEFCtrl cultured in DMEM + 10% FBS in basal condition and after sequential addition of oligomycin (O), FCCP, and antimycin A/rotenone (A/R). d, Left: western blot for IFT88 of total cell lysates from mIMCD3Ift88 and mIMCD3Ctrl generated by CRISPR/Cas9 technology. Right: Representative IF images of mIMCD3Ift88 and mIMCD3Ctrl. Cilia (ARL13B, green), nuclei (DAPI, blue). Scale bar: 10 µm. Arrows indicate cells in the insets. e, Analysis of OCR measurement in one representative experiment in mIMCD3Ift88 and mIMCD3Ctrl cultured in DMEM/F12 + 10% FBS in basal condition and after sequential addition of O, FCCP, and A/R. f, Left: analysis of OCR measurement in one representative experiment in MEFIft88 and MEFCtrl cultured in DMEM + 0% FBS in basal condition and after sequential addition of O, FCCP, and A/R. Right: analysis of OCR measurement in one representative experiment in mIMCD3Ift88 and mIMCD3Ctrl cultured in DMEM/F12 + 0% FBS in basal condition and after sequential addition of O, FCCP, and A/R. g, Principal Component analysis (PCA) of extracellular metabolites assessed by NMR spectroscopy as in Fig. 1c. h, Hierarchical clustering of extracellular metabolites assessed by NMR spectroscopy as in Fig. 1c, including metabolite concentrations in unconditioned medium (MO) and media conditioned by MEFIft88 (KO) and MEFCtrl (CT) cells. Data in dot and bar plots are mean ± SD.
