Fig 2: A hypothesis model depicting that TKI treatment increased the expression of TRIM15 by inhibiting the AKT/FOXO1 axis in HCC.The upregulated TRIM15 contributed to TKI resistance by promoting the ubiquitination and nuclear localization of LASP1, which activated the AKT signaling pathway and increased the snail expression in HCC.

Fig 3: The methylation of all CpGs in the promoter regions of five genes(A–E) Promoter region methylation of TBX4 (A), TRIM15 (B), C6orf201 (C), ARHGEF4 (D), and OR4D11 (E) in the training set (left), test cohort 1 (medium), and test cohort 2 (right). Data were presented as mean ± SEM. Student’s t test was used. ns, not significant; *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001.

Fig 4: TRIM15 interacts with LASP1 to regulate sensitivity to TKIs in HCC.a, b The whole cell lysate of Huh7 and Hep3B were collected. The Co-IP was performed by using the IgG, TRIM15 or LASP1 antibodies. c The HA-LASP1 plasmids were transfected into the Huh7 cells. After 48 h, cells were harvested and subjected to GST-pull down assay. d Huh7 and Hep3B cells were infected with indicated shRNAs for 72 h. Cells were harvested for western blot analysis. e Huh7 and Hep3B cells were infected with indicated shRNAs. After 48 h, cells were transfected with plasmids for 24 h. Cells were harvested for western blot analysis. f Huh7 and Hep3B cells were infected with indicated shRNAs for 72 h. Cells were harvested and treated with a serial dose of sorafenib for 24 h. Then, these cells were collected for CCK-8 assay. N = 3, ***, P < 0.001. g–i Huh7 and Hep3B cells were infected with indicated shRNAs. After 72 h, cells were harvested for transwell assay (g), angiogenesis assay (h), and migration assay (i). Data presents as mean ± SEM with three replicates. Ns not significant; *, P < 0.05; ***, P < 0.001. j–m Huh7 cells were infected with indicated shRNAs for 72 h. After puromycin selection, cells were collected and treated with or without sorafenib (3 µM sorafenib for cells, 50 mg/kg oral administration for nude mice) for CCK-8 assay or nude mice xenograft assay. The tumor image was shown in panel (k), the tumor mass in panel (l), and the growth curve in panel (m). For CCK-8 assay, data presents as mean ± SEM with three replicates. Ns not significant; ***, P < 0.001. For xenograft assay, data presents as mean ± SEM with three replicates. Ns not significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Fig 5: The protein levels of TBX4, TRIM15, C6orf201, and ARHGEF4 in LUSC(A–D) Representative images of human normal adjacent lung tissues and LUSC tissues stained with TBX4 (A), TRIM15 (B), C6orf201 (C), or ARHGEF4 (D) antibodies as indicated. Scale bar, 20 µm.(E) Immunohistochemical (IHC) staining of 4 antibodies in 10 pairs of normal and LUSC tissues was quantified by mean IOD. N, normal; C, cancer. Scale bar, 20 µm. Data were presented as mean ± SEM. Paired Student’s t test was used. ***, p < 0.001.