Fig. 4
Fbxl6 elevation synergizes with KrasG12D to drive HCC by upregulating Prelid2. a Venn diagram showing genes that were significantly differentially expressed at the mRNA level between liver tissues from KC and KLC mice and those from LC and KLC mice. b Heatmap showing the top 21 gene signatures representing different groups. c qPCR was used to detect the mRNA levels of Prelid2, Slc41a3 and Gldn in WT, LC, KC and KLC mice. d HCC tissues were collected from LC, KC and KLC mice for HE and IHC staining. Representative consecutive IHC staining images for Prelid2, Slc41a3 and Gldn are presented. Scale bars = 200 or 50 μm. e Cells that were positive for Prelid2, Slc41a3, or Gldn were counted among a total of 500 cells on average from 3 independent tumors derived from 3 mice per group. f The Prelid2 knockdown efficiency was determined by qPCR. Knockdown of Prelid2 inhibited cell proliferation (g), migration (h), and ROS generation (i) in KLC primary cells. j Blockade of ROS with NAC (1 or 10 mmol/L) inhibited cell migration in KLC primary cells. Scale bars = 250 (h), 200 (i), or 100 (j) μm. Unpaired Student’s t test was used to analyze the data in (i). One-way ANOVA with Tukey’s multiple comparisons test was used to analyze the data in (c, e, f, j). *P < 0.05; **P < 0.01; ***P < 0.001. Fbxl6 F-box and leucine-rich repeat 6, Kras kirsten rat sarcoma, Prelid2 the proteins of relevant evolutionary and lymphoid interest (PRELI) domain 2, WT wild-type, LC LSL-Fbxl6KI/+;Alb-Cre, KC LSL-KrasG12D/+;Alb-Cre, KLC LSL-KrasG12D/+;LSL-Fbxl6KI/+;Alb-Cre, Slc41a3 solute carrier family 41 member 3, Gldn gliomedin, IHC immunohistochemistry, HCC hepatocellular carcinoma, ROS reactive oxygen species, NAC N-acetylcysteine