Fig. 5: Primary cilia promote EMT and metastasis upon Gln deficiency.

A, B Primary cilia formation was observed in -QQ PANC-1 cells. (A) Primary cilia were detected by immunofluorescence staining with an antibody against ARL13b (green) in Prt or -QQ PANC-1 cells. DNA was stained with DAPI (blue). Scale bar, 20 µm. B Quantitative results of the frequency of ciliated cells in (A). C Gln supplementation inhibited primary ciliogenesis in -QQ cells. Quantitative results of the frequency of ciliated cells supplemented with different dosages of Gln in -QQ PANC-1 cells. Ciliary components were detected by immunofluorescence staining with antibodies against acetylated tubulin (Ac-tub, D–F), CEP164 (D), ARL13b (E), and IFT88 (F). DNA was stained with DAPI. Scale bar, 10 µm. G–I Primary cilia contributed to EMT and cell invasion. (G) IFT88 was depleted efficiently. Extracts of parental (Prt) or -QQ PANC-1 cells in the absence or presence of siRNA against IFT88 (siIFT88) were analyzed by western blot assay with antibodies against IFT88, E-cadherin (E-cad), or Ku70. (H) Quantitative results of the frequency of ciliated cells in Prt or -QQ PANC-1 cells in the absence or presence of siRNA against IFT88 (siIFT88). I Quantitative results of the relative invaded cells in Prt or -QQ PANC-1 cells in the presence or absence of siRNA against IFT88 (siIFT88). J The frequency and length of primary cilia were significantly increased in the -QQ PDAC spheroids. Primary cilia were detected by immunofluorescence staining with an antibody against ARL13b (white) in Prt or -QQ PANC-1 spheroids. DNA was stained with DAPI (blue). Scale bar, 10 µm. Data are represented as the mean ± SD of three independent experiments. n.s. no significance, ***P < 0.001.