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. 2013 Apr 8;110(17):6829–6834. doi: 10.1073/pnas.1217002110

Fig. 4.

Fig. 4.

Y828 phosphorylation is required for CD133 maintaining glioma stem cell self-renewal and tumorigenesis. (A and B) Single-cell neurosphere formation assay of CD133+ cells expressing LacZ shRNA, CD133 shRNA1, CD133 shRNA1+shRNA-resistant WT CD133, or CD133 shRNA1+shRNA-resistant Y828F. (A) Representative images of neurospheres are shown. (B) The percentage of neurosphere-containing wells in each group. (Scale bar, 10 µm.) (C and D) A total of 1, 10, or 100 CD133+ cells isolated from T21109 (C) or T12179 (D) samples infected with the indicated lentivirus were cultured in 96-well plates. After 10 d, the number of neurosphere was counted. Results are expressed as mean ± SD from three separate experiments; ***P < 0.001. (E) WT CD133, not Y828F mutant, rescues the effect of CD133 knockdown on the tumor-initiating capacity of CD133+ cells. An intracranial limiting dilution tumor formation assay (using 10,000, 5,000, 1,000, and 500 cells per mouse) was performed using CD133+ cells infected with the indicated lentivirus. The table displays the number of mice developing tumors.