![ARF3 regulates metastasis in vivo. (A) Schema, intraprostatic transplantation of PC3 cells expressing mNG or ARF3-mNG and Scr or ARF3 shRNA into CD-1 nude male mice. PT formation and incidence and location of MM determined after 8 wk. (B) PT incidence in mice (total number) transplanted with PC3 cells expressing mNG, Scr shRNA (control, 20 mice), mNG, ARF3 shRNA (18 mice), or ARF3-mNG, Scr shRNA (17 mice). P values (Chi-squared test), indicated. (C) Prostate weight in mice (with PT only) transplanted with PC3 cells expressing mNG, Scr shRNA (control, 12 mice), mNG, ARF3 shRNA (12 mice, weight of 1 mouse prostate not recorded), or ARF3-mNG, Scr shRNA (9 mice). Box and whiskers plot, min–max percentile; +, mean; dots, outliers; midline, median; boundaries, quartiles. P values (Student’s two-tailed t test). (D) MM incidence in mice (with PT only) transplanted with PC3 cells expressing mNG, Scr shRNA (control, 12 mice), mNG, ARF3 shRNA (12 mice), or ARF3-mNG, Scr shRNA (9 mice). P values (Chi-squared test), indicated. (E) MM count/mouse in mice (with PTs only) transplanted with PC3 cells expressing mNG, Scr shRNA (control, 12 mice), mNG, ARF3 shRNA (12 mice), or ARF3-mNG, Scr shRNA (9 mice). Box and whiskers plot, min–max percentile; +, mean; dots, outliers; midline, median; boundaries, quartiles. P values (Mann–Whitney test [two-tailed]), indicated on graph. (F) MM frequency to indicated organs for mice (with PT only) transplanted with PC3 cells expressing mNG, Scr shRNA (control, 12 mice), mNG, ARF3 shRNA (12 mice), or ARF3-mNG, Scr shRNA (9 mice). FP, fat pad. (G) Serial sections representative of a primary tumor from a mouse transplanted with PC3 cells expressing mNG, Scr shRNA, mNG, ARF3 shRNA, or ARF3-mNG, Scr shRNA, stained for H&E and N-cadherin (left panels and higher magnification). Scale bars, 1 mm or 100 μm. (H and I) Percentage of N-cadherin–positive PT (H) or weighted Histoscore for N-cadherin in PTs (I) in 8 mNG Scr shRNA, 7 mNG ARF3 shRNA, and 5 ARF3-mNG, Scr shRNA mice. Box and whiskers plot, 10–90 percentile; +, mean; dots, outliers; midline, median; boundaries, quartiles.](https://cdn.rupress.org/rup/content_public/journal/jcb/222/4/10.1083_jcb.202206115/2/jcb_202206115_fig8.png?Expires=1771055109&Signature=MnCbkPa6nyPV9nWUJ4ZSQs7ceic5py3p8yQMnVWneIJtb3BNyEDUkhhGaXHWUQx7ATI~cAPpmy~yzLP3JcJmXcGB8QyupFgMhhzox0HKGmSEJ~W9k-PXd1jQIeDOGCR8IgMDjgfzqtedhJLOxHUUFx59vExJWmRMru5RwowCGI3Qtx3utFBxC5r6gScNiZbqpwSmkp5tXBz6XdUdY3J2OTM0znvK3shwXCOXzmZq0bPKLCVJFFc6ID4ikmshbvpCX~3ttO1hez2SfEKNigEJLAUUny6PA~AhMPs-x~R5Iyx-L-8tdkUdMEX1ImuNWObwPPPLh2pmjateP~0ii5wf8g__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
ARF3 regulates metastasis in vivo. (A) Schema, intraprostatic transplantation of PC3 cells expressing mNG or ARF3-mNG and Scr or ARF3 shRNA into CD-1 nude male mice. PT formation and incidence and location of MM determined after 8 wk. (B) PT incidence in mice (total number) transplanted with PC3 cells expressing mNG, Scr shRNA (control, 20 mice), mNG, ARF3 shRNA (18 mice), or ARF3-mNG, Scr shRNA (17 mice). P values (Chi-squared test), indicated. (C) Prostate weight in mice (with PT only) transplanted with PC3 cells expressing mNG, Scr shRNA (control, 12 mice), mNG, ARF3 shRNA (12 mice, weight of 1 mouse prostate not recorded), or ARF3-mNG, Scr shRNA (9 mice). Box and whiskers plot, min–max percentile; +, mean; dots, outliers; midline, median; boundaries, quartiles. P values (Student’s two-tailed t test). (D) MM incidence in mice (with PT only) transplanted with PC3 cells expressing mNG, Scr shRNA (control, 12 mice), mNG, ARF3 shRNA (12 mice), or ARF3-mNG, Scr shRNA (9 mice). P values (Chi-squared test), indicated. (E) MM count/mouse in mice (with PTs only) transplanted with PC3 cells expressing mNG, Scr shRNA (control, 12 mice), mNG, ARF3 shRNA (12 mice), or ARF3-mNG, Scr shRNA (9 mice). Box and whiskers plot, min–max percentile; +, mean; dots, outliers; midline, median; boundaries, quartiles. P values (Mann–Whitney test [two-tailed]), indicated on graph. (F) MM frequency to indicated organs for mice (with PT only) transplanted with PC3 cells expressing mNG, Scr shRNA (control, 12 mice), mNG, ARF3 shRNA (12 mice), or ARF3-mNG, Scr shRNA (9 mice). FP, fat pad. (G) Serial sections representative of a primary tumor from a mouse transplanted with PC3 cells expressing mNG, Scr shRNA, mNG, ARF3 shRNA, or ARF3-mNG, Scr shRNA, stained for H&E and N-cadherin (left panels and higher magnification). Scale bars, 1 mm or 100 μm. (H and I) Percentage of N-cadherin–positive PT (H) or weighted Histoscore for N-cadherin in PTs (I) in 8 mNG Scr shRNA, 7 mNG ARF3 shRNA, and 5 ARF3-mNG, Scr shRNA mice. Box and whiskers plot, 10–90 percentile; +, mean; dots, outliers; midline, median; boundaries, quartiles.
