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. 2021 Apr 23;42(9):1486–1497. doi: 10.1038/s41401-021-00657-w

Fig. 3. Triptolide reduces β-catenin expression in NCI-H1299 and NCI-H460 cells.

Fig. 3

a–c The effect of triptolide on the protein and mRNA levels of β-catenin in NCI-H1299 and NCI-H460 cells. The cells were treated with triptolide for the indicated times and then collected for Western blotting assays and RT-qPCR assays. Representative images and quantitative Western blotting data are presented in a and b. The RT-qPCR results are shown in c. d, e The effect of triptolide on the nuclear translocation of β-catenin. The cells were lysed by using a NE-PERTM Nuclear and Cytoplasmic Extraction Kit and subjected to Western blotting assays. β-Actin and Lamin B were used as loading controls. Representative images and quantitative data are shown in d and e, respectively. f, g The effect of triptolide on β-catenin translocation from the cytoplasm to the nucleus, as indicated by immunofluorescence assays. After treatment with triptolide for 24 h, the cells were fixed with 4% paraformaldehyde, blocked using 5% BSA, and incubated with β-catenin antibody, followed by Alexa Fluor 594-conjugated donkey anti-rabbit IgG and DAPI. TPL triptolide. Representative images and quantitative data are shown in f and g, respectively. Images were taken using a Zeiss LSM 800 confocal microscope with a 63× oil immersion lens. TPL triptolide. Quantitative data are presented as the mean ± SEM, n = 3. *P < 0.05, **P < 0.01 and ***P < 0.001 vs. the vehicle group.