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. Author manuscript; available in PMC: 2020 Feb 1.
Published in final edited form as: Mol Cancer Res. 2018 Nov 14;17(2):420–430. doi: 10.1158/1541-7786.MCR-18-0455

Figure 6. Faslo colon CSC-like cells exhibit a higher lung colonization potential and resistance to T cell immunotherapy.

Figure 6.

A. CT26 cells were stained with CD133-, CD24- and Fas-specific mAbs and sorted into CD133+CD24loFaslo and CD133+CD24hiFashi cells. Showing is the gating strategy for sorting. B. The sorted CD133+CD24loFaslo and CD133+CD24hiFashi cells were injected i.v. into BALB/c mice (1.5×105 cells/mouse, n=5). Fourteen days later, mice were sacrificed and india ink was perfused into the lung. The ink-inflated lungs were fixed. Shown are tumor-bearing lungs. The tumor nodule number was counted and presented at the right. Statistical significance was determined by student t test. C. MC38.met cells were stained with CD133-, CD24- and Fas-specific mAbs and sorted into CD133+CD24loFaslo and CD133+CD24hiFashi cells. Showing is the gating strategy for sorting. D. The sorted CD133+CD24loFaslo and CD133+CD24hiFashi cells were injected i.v. into C57BL/6 mice (3×105 cells/mouse, n=5). Fourteen days later, mice were sacrificed and india ink was perfused into the lung. The ink-inflated lungs were fixed. Shown are tumor-bearing lungs. The tumor nodule number was counted and presented at the right. E. Sorted CD133+CD24loFaslo (n=6) and CD133+CD24hiFashi (n=7) MC38.met cells were injected into FasL-deficient faslgld mice (3×105 cells/mouse) i.v. Fourteen days later, mice were sacrificed and india ink was perfused into the lung. The ink-inflated lungs were fixed. Shown are tumor-bearing lungs. The tumor nodule number was counted and presented at the bottom. F. CD133+CD24loFaslo and CD133+CD24hiFashi CT26 cells were injected i.v. into BALB/c mice (2×105 cells/mouse, n=5). Four days later, mice with treated with saline control or perforin-deficient pk03 CTLs (3×105 cells/mouse). Mice were sacrificed on day 14 and analyzed as in B. G. CD133+CD24loFaslo and CD133+CD24hiFashi CT26 cells were injected i.v. into BALB/c mice (2×105 cells/mouse, n=5). Four days later, mice were treated with IgG (200 mg/mouse) or anti-PD-1 mAb (200 mg/mouse) every 2 days for 5 times. Lung tumors were analyzed as in B. H. CD133+CD24loFaslo and CD133+CD24hiFashi CT26 cells were injected s.c. into BALB/c mice (2×105 cells/mouse, n=5). Ten days later, CD133+CD24loFaslo and CD133+CD24hiFashi CT26 tumor-bearing mice were randomly grouped into two groups, respectively, and treated with IgG or anti-PD-1 mAb as in G every 2 days for 5 times. Mice were sacrificed two days after the last treatment. Shown are tumor images. I. The sizes and weights of tumors as shown in H were measured and analyzed by student t test.