Fig. 6: Plekha5 is a tumor metastasis suppressor.

a Representative IHC images of Plekha5 in 153PT and 153LMT. Scale bar, 50 µm. b Representative IHC images of Plekha5 in primary tumor formed by Plekha5-KO B477-GFP cells injected into the mammary fat pad of nude mice. Scale bar, 50 µm. c Transwell assays using Plekha5-KO B477-GFP cells. Scale bar, 1 mm (n = 3 replicates over three independent experiments). Data are reported as the mean ± SD. Significance determined by two-tailed Student’s t test. d Metastasis functional assay in vivo using Plekha5-KO B477-GFP cells. Top: wholemount lungs or livers from control mice. Bottom: wholemount or frozen section of livers from mice implanted with Plekha5-KO cells. Left four, merged image of left two and three with 4′,6-diamidino-2-phenylindole (DAPI). Representative images were shown. Scale bar: wholemount, 3 mm; frozen section: 20 µm. BF, bright field. e Quantification of GFP signals in organs of mice (n (control group) = 9 mice, n (Plekha5-KO group) = 10 mice). Count the number of more than 10 as 10. Significance determined by two-tailed Student’s t test. f Representative IHC images of PLEKHA5 in primary tumor formed by PLEKHA5-KO MDA-MB-231-GFP cells injected into the mammary fat pad of nude mice. Scale bar: 50 µm. g Transwell assays for PLEKHA5-KO MDA-MB-231-GFP cells. Scale bar: 1 mm (n = 3 replicates over three independent experiments). Data are reported as the mean ± SD. Significance determined by two-tailed Student’s t test. h Metastasis functional assay in vivo using PLEKHA5-KO MDA-MB-231-GFP cells. Top: wholemount (scale bar, 3 mm) and enlarged (scale bar, 0.3 mm) images of lungs from control mice. Bottom: wholemount (scale bar, 3 mm) and enlarged (Scale bar, 0.3 mm) images of lungs from mice implanted with PLEKHA5-KO cells. i Quantification of GFP signals in organs of mice (n (control) = 10 mice, n (PLEKHA5-KO) = 11 mice. Significance determined by two-tailed Student’s t test. j Metastasis incidence of mice after implanted with different Plekha5/PLEKHA5-KO mouse or human cell lines. k Expression of PLEKHA5 in the primary tumors without metastasis (n = 107 tumors) and the primary tumors with metastasis (n = 48 tumors). Significance determined by two-tailed Student’s t test. Box-and-whisker plots: center line, median; box limits, upper and lower quartiles; whiskers, minima to maxima. l Kaplan–Meier curves showing the correlation between PLEKHA5 expression and overall survival. Significance determined by a two-sided the log-rank test (Mantel–Cox). Error bars represent SD.