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. 2021 Nov 23;12:752825. doi: 10.3389/fphar.2021.752825

FIGURE 5.

FIGURE 5

DSF/Cu induced autophagic cell death by upregulating ULK1. (A) CQ can partially reverse the cell viability caused by the DSF/Cu treatment in RKO (p < .01). (B,C) Low expression of ULK1 in CRC cell lines is established in RKO and Ht2, and the efficiency was verified by Western blot and qRT-PCR. (D,E) CCK8 assay showed that cell viability could be partially reversed in the low ULK1 expression group after incubation with DSF/Cu (DSF 0–0.2 μM/Cu 10 μM in RKO cells, DSF 0–0.3 μM/Cu 10 μM in HT29 cells) for 48 h (p < .01). (F) Western blot showed that DSF/Cu combined with CQ or Bafa1 could not enhance the accumulation of LC3-II in shULK1-RKO. (G)Colony formation assay showed that cell viability could be partially reversed in the low ULK1 expression group after incubation with DSF/Cu (0.25/10 μM) for 48 h (p < .01). (H,I) The expression of LC3 detected by immunofluorescence in the shULK1-RKO group was lower than the shNC-RKO group after incubation with DSF/Cu (0.25/10 μM) for 48 h (p < .01).