Fig 2: The overexpression of SAE1 is associated with metastasis and poor prognosis in patients with HCC. (A) Representative IHC photo-images and graphical representation of SAE1 protein level in paracancerous and cancerous tissues. (**** p < 0.0001) (B) Kaplan-Meier curve of overall survival according to SAE1protein level of TMU-SHH HCC cohort. (C) Representative IHC photo-images of SUMO1, SUMO2, and UBC9 protein levels in paracancerous and cancerous tissues.

Fig 3: miR-382-3p represses SAE1 expression directly by targeting its 3'UTR. (A) Dual-luciferase assay. (B) miR-382-3p mimics or inhibitor were transfected into A549 and H1299 cells. At 48 h after transfection, the cells were subjected to protein exaction and immunoblotting. (C) The levels of SAE1 and Ki-67 in xenograft tumors generated from miR-382-3p-overexpressing A549 and H1299 cells were examined by immunohistochemistry (scale bars, 50 µM). *P<0.05, **P<0.01. SAE1, small ubiquitin-like modifier 1 activating enzyme subunit 1; miR, microRNA; WT, wild-type; Mut, mutant.

Fig 4: Expression of SAE1, a target of miR-382-3p, is negatively correlated with miR-382-3p levels in tumors. (A) Gene Ontology analysis of predicted miR-382-3p targets. (B and C) SAE1 protein levels in tumor and adjacent non-tumor control samples were examined by immunoblotting. (B) Representative western blots and (C) comparison of SAE1 protein levels between tumor and adjacent non-tumor control samples for each case. (D) Correlation analysis of the SAE1 protein levels and miR-382-3p levels in lung adenocarcinoma tumor samples. (E) Kaplan-Meier curves depicting the overall survival of 1,044 patients with lung cancer. Data were from Kaplan-Meier Plotter. SAE1, small ubiquitin-like modifier 1 activating enzyme subunit 1; miR, microRNA; MF, molecular function; CC, cellular component; BP, biological process; T, tumor tissue; N, normal tissue.

Fig 5: miR-382-3p inhibition promotes proliferation and inhibits apoptosis in lung adenocarcinoma cell lines. A549 and H1299 cells were transfected with miR-382-3p inhibitor, with or without SAE1-specific siRNA for 48 h. (A) The cells were subjected to MTT assays to examine proliferation and viability. (B) Flow cytometric analysis was used to detect cell viability and apoptosis. *P<0.05, **P<0.01. (C and D) Cell cycle analysis of (C) A549 and (D) H1299 cells. *P<0.05 S phase vs. inhibitor control; #P<0.05, ##P<0.01 G2-M phase vs. inhibitor control. ns, no significance; miR, microRNA; PI, propidium iodide; APC, allophycocyanin; siRNA, small interfering RNA; si-SAE1, siRNA targeting SAE1; SAE1, small ubiquitin-like modifier 1 activating enzyme subunit 1; FL-H, fluorescence-height.