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. 2023 Jul 24;8(14):e167310. doi: 10.1172/jci.insight.167310

Figure 6. Pharmacologic inhibition of Notch signaling reduces LIN28B-induced liver metastasis.

Figure 6

(A) qRT-PCR analysis of NOTCH3 mRNA in DLD-1 LIN28Bhi cells treated with DMSO, 10 μM DAPT, or 20 μM DAPT. (B) qRT-PCR analysis of HES1 mRNA in DLD-1 LIN28Bhi cells treated with DMSO, 10 μM DAPT, or 20 μM DAPT. (C) Representative images of the wound closure scratch assay performed using DLD-1 LIN28Bhi cells treated with DMSO, 10 μM DAPT, or 20 μM DAPT. Scale bar = 200 μm. Area of the wound was measured by using ImageJ. Data represented as means ± SEM (n = 3). (D) Representative images of the aggregation assay performed using DLD-1 LIN28Bhi cells treated with DMSO, 10 μM DAPT, or 20 μM DAPT. The number of cell aggregates was counted using Keyence BZ-X810. Data represented as means ± SEM (n = 3). (E) Schematic drawing of the workflow used to study the effects of DAPT injection in a portal vein injection model of CRC metastasis. (F) Representative images of RFP expressed by DLD-1 LIN28Bhi tumors in the liver. Images were taken using Keyence BZ-X810. Scale bar = 500 μm. Data in A–D were analyzed by 1-way ANOVA followed by Tukey’s multiple-comparison test. Data in F were analyzed by Fisher’s exact test. **P < 0.01, ***P < 0.001, ****P < 0.0001.