Fig. 3

CRART16 promotes cetuximab resistance and contributes to the acquisition of stemness properties of CRC cells. a CCK8 assay was performed to assess the cell viability of Caco-2-CRART16 and Caco-2-NC cells treated with graded concentrations of cetuximab (0–200 μg/ml) for 48 h. Each concentration had six replicates, and the experiment was repeated at least three times. Data are presented as mean ± SD. **P < 0.01 by two-way ANOVA. b Flow cytometry was performed in Caco-2-CRART16 and Caco-2-NC cells with cetuximab treatment (100 μg/ml and 200 μg/ml) for 48 h. All experiments were repeated three times, and data are presented as mean ± SD. **^/##P < 0.01 by Student’s t test. c Caco-2-CRART16 and Caco-2-NC cells were treated with 200 μg/ml cetuximab for 48 h. Apoptosis was detected by a TUNEL assay. Scale bar = 100 μm. d The cell cycle was assessed by flow cytometry in Caco-2-CRART16 and Caco-2-NC cells after 48 h of treatment with cetuximab (200 μg/ml). All experiments were repeated three times, and data are presented as mean ± SD. *P < 0.05, **^/##P < 0.01 by Student’s t test. e The percentage of EGFR-, ERBB3-, and c-MET-positive cells and the MFI were determined by a GALLIOUS flow cytometer in Caco-2-CRART16 and Caco-2-NC cells. All experiments were repeated three times, and data are presented as mean ± SD. **P < 0.01 by Student’s t test. f Flow cytometry analysis showed the expression of stemness biomarkers in CRC cells, CD44 and CD133, in Caco-2-CRART16 and Caco-2-NC cells. All experiments were repeated three times, and data are presented as mean ± SD. *P < 0.05 by Student’s t test