Fig 1: Combination treatment of etravirine and paclitaxel suppressed tumor growth and metastasis in the A2780 orthotopic xenograft model. (a,b) A2780 cells were injected into the ovarian bursa sac of nude mice (five mice per group). Randomized mice were treated with control (saline), etravirine (100 mg/kg x three times per week by i.p. injections), paclitaxel (15 mg/kg once a week), and a combination group for three weeks. Representative fluorescence images of female nude mice after three weeks of treatment. (c,d) Tumor weight and tumor size were measured at the end of the treatment. (e) Mice body weights were monitored weekly. (f,g) Metastatic tumors were counted and measured. (h) Representative H&E staining, IHC images, and quantification of cell proliferation marker PCNA, AGR2, angiogenesis marker CD31, and apoptosis marker cleaved caspase-3. Data are presented as mean ± SEM. One-way ANOVA and t-test analyses. * p < 0.05, ** p < 0.01, *** p < 0.001.
Fig 2: Etravirine induces AGR2 degradation by autophagy. (a–d) Western blot analysis showed the effect of etravirine on AGR2 expression and LC3B expression. (e,f) Immunofluorescence analysis showed the expressions of AGR2 and LC3B in A2780-ADR cells after treatment with 5 µM etravirine for 48 h and 50 µM CQ for 24 h by using Lion Heart FX automated microscope (40×). Scale bar 100 µm. (g,h) A2780 and A2780-ADR cells were exposed with wortmannin (PI3K inhibitor) 1 µM for 24 h prior to etravirine 10 µM for 72 h. (i,k) A2780 and A2780-ADR cells were pretreated with 50 µM CQ for 24 h and then treated with 10 µM etravirine for 72 h. (l) Knockdown of ATG7 by 100 nM siRNA for 72 h in A2780 cells and then treated with etravirine 10 uM for 72 h, AGR2 and LC3B expression were detected by Western blot. Data are shown as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001.
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