Fig 2: Prediction and identification of the ANXA10 ceRNA network. (A) Venn diagram displaying ANXA10-targeting miRNAs in the PITA, miRanda, and TargetScan tools. (B) Significant correlations of ANXA10 and targeted miRNAs analyzed by starBase. (C) Venn diagram showing the targeting-lncRNAs of hsa-miR-758-3p predicted by starBase and miRNet2.0. (D) Scatter plots displaying the close correlations between hsa-miR-758-3p expression and ENTPD3-AS1, LAMTOR5-AS1 and LINC01278 expression. (E) Sankey diagram showing the correlations in the final ceRNA network.

Fig 3: UV-crosslinking competition experiments employing rat Anxs and radiolabeled [α32P]-rUTP c-myc or anxA2 3′UTRs. 2 μM purified rat Anxs (except 0.4 µM of AnxA7; 0.3 µM of AnxA9 and 0.5 µM of AnxA11) were UV-crosslinked in the absence of RNA (lanes 1 and 5). 100,000 cpm of radiolabeled anxA2 3′UTR (7 fmoles) or c-myc 3′UTR (6 fmoles) were UV-crosslinked to purified rat Anxs in the absence (lanes 2 and 6) or presence of 25x (lanes 3 and 7), or 50x (lanes 4 and 8) molar excess of the corresponding unlabeled transcript. 2 μM BSA and mutant AnxA2 served as negative controls. After UV-crosslinking and RNase treatment, the samples were subjected to 4%–15% SDS-PAGE and the proteins were stained with Coomassie Brilliant Blue (lanes 1–4), whereafter the gels were dried. The [α32P]-rUTP-labeled RNA covalently bound to the respective Anxs, as indicated, was visualized using screens and phosphor-imaging following an overnight (c-myc 3′UTR) or 6 h (anxA2 3′UTR) exposure (lanes 5–8). PageRuler prestained protein ladder (from top to bottom: 100 kDa, 70 kDa (the most prominent band), 55 kDa, 40 kDa, 35 kDa and 25 kDa) are shown to the left of the AnxA1, AnxA3, AnxA5, AnxA7, AnxA8, AnxA10, Δ188AnxA11 and BSA samples. The representative images are from two experiments performed with competition while RNA-Anx binding was performed four times without competition.

Fig 4: Correlation of ANXA10 expression with m6A genes in TCGA-LIHC. The m6A genes discrepancy of expression in the low and high ANXA10 expression groups with TCGA (A) and GSE14520 (B) data. (C) Kaplan–Meier curve of HNRNPA2B1, IGF2BP3, METTL3, RBM15, YTHDF1 and YTHDF2 in LIHC after TCGA-LIHC data and GSE14520 data were merged. *p < 0.05; **p < 0.01; ***p < 0.001.

Fig 5: Correlation and analysis of immunomodulators related to ANXA10. (A) Correlation heatmaps showed that ANXA10 expression was associated with immunoinhibitors (left) and immunostimulators (right). (B,C) GO and KEGG analyses of ANXA10-related immunomodulatory genes via WebGestalt.