Fig. 1: A phenotypic transition of GBM cells upon nutritional stress. | Cell Death Discovery

Fig. 1: A phenotypic transition of GBM cells upon nutritional stress.

From: Nutritional stress reprograms dedifferention in glioblastoma multiforme driven by PTEN/Wnt/Hedgehog axis: a stochastic model of cancer stem cells

Fig. 1

a Schematic representation of the workflow of gradual nutrient depletion. Single-cell suspensions were made and cells (5 × 105) were seeded in a six-well plate in 2 ml of IMDM with 10% FBS and cultured for 5 days in the CO2 incubator without replenishment of fresh medium. b Representative phase-contrast images with ×10 magnification, showing the sphere-like appearance of GBM cells (iU87MG) upon 5 days of nutritional stress, whereas there was no sphere formation upon continually changing the medium. c Shifting of cell sizes toward smaller-sized population (gated as R2) of a fraction of the remaining population (gated as R1) at day zero (D0), 2 (D2), and 5 (D5) as assessed by flow cytometry. d Graphical representation of the percentage of cells shifting toward smaller-sized R2 population at D0, D2, and D5. e Propidium iodide (PI) staining showed a very low percentage of apoptotic cells in the R1 and R2 population after 5 days of nutritional stress. f Representative histogram plots showing differential expression of GSC markers (CD133-APC, CD90-PECy5, and CD117-PE) in the R1 and R2 population as represented by mean fluorescence intensity (MFI) at day 5 of nutritional stress. White peaks represent autofluorescence of the cells in the R1 and R2 populations, and gray peaks represent MFI of GSCs markers (CD133-APC, CD90-PECy5, and CD117-PE) in the R1 and R2 populations. Values on the peak represent the MFI values of the markers. g Cell surface staining followed by flow cytometry analysis determined the percentage of CD133-APC-, CD90-PECy5-, and CD117-PE-positive cells in the R1 and R2 population at day 2 and 5 of nutritional stress. h Cell surface staining at different time points (0, 2, and 5 days) of nutritional stress followed by flow cytometry analysis determined time-dependent enhancement in the expression of GSCs markers in the R1 population. i Flow cytometric analysis revealed time-dependent stochastic increase of the number of CD133-APC-, CD90-PECy5-, and CD117-PE-positive cells in the total population under nutritional stress. j Flow cytometric analysis showed a slight increase in the MFI of GSCs makers in the total population at different time points due to nutritional stress. The results represent mean ± SD from three independent experiments

Back to article page