Fig. 4
RNF43 enhanced lung adenocarcinoma cell metastasis through EMT. a, Phase contrast microscopy of A549 + control shRNA cells (Control) or RNF43 shRNA cells (RNF43 KD) plated on dishes over a 5-day period. Scale bars, 50 μm. b, Ectopically grown A549 cells were surgically removed. Biweekly quantification of bioluminescence showed accelerated tumor growth and increased spontaneous metastasis in the mice implanted with A549 + Control cells relative to that in mice implanted with A549 + RNF43 KD cells (t test, *, P < 0.05). Data are presented as the mean ± SD. c, Representative bioluminescence images of tumor cells before and after primary tumor resection (10 and 30 days after resection). d, HE staining of micrometastasis to bone, lung, liver, and brain tissue. Scale bars, 100 mm. e, Total cumulative incidences of micrometastasis in the tumor-implanted mouse cohorts as confirmed by immunostaining (χ2 test, *, P < 0.05). Micrometastatic events in various BALB/c nude mouse organs were confirmed by immunostaining. Tissue sections were scored as positive or negative based on the presence or absence of detectable micrometastasis. f, Kaplan-Meier survival analysis of the corresponding mice showed significantly shorter survival times for mice implanted with A549 + Control cells compared with that of mice implanted with A549 + RNF43 KD cells (log-rank test, **, P < 0.01)