Fig. 6
Anti-TM4SF5 isoxazoles recovered SLAMF7-positive cell populations to block cellular growth in an HCC model. a Scheme for an HCC PDX xenograft model in NOD-SCID male mice (6-week-old, n = 8). b Tumors were extracted from the right flank of mice 36 days after tumor inoculation. c Tumor size was measured twice a week with a caliper. Tumor weight was measured after sacrifice and tumor extraction. d Tumor tissues were immunostained for the indicated molecules. Representative images are shown. e, f HCC patient tissues [two TM4SF5-positive cases of #7 and #17 (red); two TM4SF5-negative cases of #12 and #15 (blue) and one case of #8 without increased TM4SF5 (gray)] were immunoblotted for TM4SF5 and SLAMG7 (e). Whole tissue extracts were processed for immunoprecipitation using anti-LAMP1 antibody before immunoblotting for the molecules (f). The ratios of normalized band intensities were compared between conditions. g Experimental protocol for an HCC model with liver-orthotopic SNU449T7 cell implantation into male BALB/c-Nude mice (n = 9). h Body weight and maximal tumor volume changes during the liver-orthotopic SNU449T7 xenograft experiments. i Slamf7-immunostained liver images of tumors from mice with vehicle or ST-5-002 treatment. j Liver tissues were analyzed for immunohistochemistry using antibodies against NK cell ligands. Two Slamf7 images in each experimental condition were at two different resolutions. Scale bars, 100 μm. *P < 0.05, **P < 0.01, ns = non-significant, unpaired Student’s t test or Two-way ANOVA, Tukey’s multiple comparisons test. Data are represented as the mean ± SEM. See also Figs. S7 and S8
