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. Author manuscript; available in PMC: 2022 Apr 1.
Published in final edited form as: Mol Cancer Res. 2021 Jun 22;19(10):1651–1665. doi: 10.1158/1541-7786.MCR-20-0994

Figure 2. Nestin deficiency induces G2/M phase arrest of GBM cells.

Figure 2

a, Nestin protein levels in U251 cells transduced with a lentivirus carrying shRNAs specific for Nestin (sh-Nes#1 and sh-Nes#2) or scrambled shRNA (shCtrl), examined by western blotting.

b–c, Images of colony formation of Nestin-deficient U251 cells (sh-Nes#1, sh-Nes#2) and control cells (transduced with scrambled shRNA) (b). The number of colonies from Nestin-deficient U251 cells and control cells was quantified (c).

d–e, Cell cycle distribution of U251 cells infected with Nestin shRNAs or scrambled shRNA, examined by flow cytometry (d). Percentages of cells in M phase among Nestin-deficient and control U251 cells was quantified (e).

f–g, Flow cytometry of U251 cells after labelling with Annexin V and PI (f). The percentage of apoptotic cells (Annexin V+), including cells at an early stage of apoptosis (Annexin V+/PI−) as well as cells at a late stage of apoptosis (Annexin V+/PI+), was quantified (g).

h–l, Nestin protein levels in tumor cells from GBM PDX1 and PDX2 lines, after viral infection with Nestin shRNA (sh-Nes#1) or scrambled shRNA, examined by western blotting. Colony formation of tumor cells from PDX-1 pr PDX-2, after infection with Nestin shRNA (sh-Nes#1 or sh-Nes#2) or scrambled shRNA (i). The number of colonies from Nestin deficient tumor cells and control tumor cells was quantified (j). Cell cycle distribution of Nestin-deficient or control tumor cells from PDX-1 (k). The percentage of PDX1 cells and PDX2 cells in G2/M phase, after infection with Nestin shRNAs or scramble shRNA, was quantified (l).