Fig 2: SNHG22 promotes Wnt/β-catenin pathway activation via HMGA1 upregulation by binding to miR-361-3p in GC cells. (A) Potential binding sites of miR-361-3p to HMGA1. (B) HMGA1 expression in GC patients predicted in GEPIA database. (C) HMGA1 expression in GC and adjacent normal tissues of GC patients detected by RT-qPCR. (D) Correlation between HMGA1 and SNHG22 expression in GC tissues (r = 0.759). (E) The correlation between HMGA1 and miR-361-3p expression in GC tissues (r = −0.775). (F) RT-qPCR to measure the effect of miR-361-3p mimic on HMGA1 mRNA expression. (G) Western blot analysis of the effect of miR-361-3p mimic on HMGA1 protein expression. (H) The effect of miR-361-3p mimic on the luciferase activity of HMGA1-WT/HMGA1-MT detected by dual-luciferase assay. (I) RT-qPCR detection on the effect of sh-SNHG22 and miR-361-3p inhibitor on HMGA1 mRNA expression. (J) Western blot analysis to assess the effect of sh-SNHG22 and miR-361-3p inhibitor on HMGA1 and β-catenin protein expression. In panel (A), **p < 0.01 according to two-way ANOVA; in panel (D) and (E), Pearce correlation analysis was conducted, p < 0.001; in panel (F–H) #p < 0.05 according to two-way ANOVA; in panel (I) and (J), and p < 0.05 vs MKN-45 and AGS cells transfected with sh-SNHG22, & p < 0.05 vs GES-1, MKN-45 and AGS cells transfected with sh-NC, & p<0.05 vs GES-1, AGS and MKN-45 cells transfected with sh-NC according to two-way ANOVA, @ p < 0.05 vs GES-1, MKN-45 and AGS cells transfected with sh-SNHG22 + NC inhibitor according to two-way ANOVA. All experiments were repeated three times independently, and the results were averaged.

Fig 3: Attenuation of HMGA1 by IL-24wt inhibits AKT activation in H1299-IL24 lung cancer cellsA. Western blotting analysis showed that expression of IL-24wt protein in H1299-IL24 cells reduced the expression of phosphorylated (p) AKTT308 and pAKTS473, and increased PPP2R2A expression at 24 h and 48 h after doxycycline treatment. B. Western blotting analysis showed that expression of IL-24wt protein in HCC827 and A549 cells reduced the expression of phosphorylated (p) AKTT308 and pAKTS473, and increased PPP2R2A expression. Beta-actin was used as a protein loading control. Differences in the expression of the proteins were determined by semi-quantitative analysis and represented in graphical format (p<0.05). Bars denote standard deviation (SD).

Fig 4: IL-24wt in combination with HMGA1 siRNA treatment exhibited enhanced inhibition of tumor cell migration and invasionsi-RNA-mediated HMGA1 knockdown combined with IL-24wt resulted in significant reduction of tumor A. cell migration and B. invasion compared with controls. IL-24 treatment alone and HMGA1 siRNA treatment alone significantly inhibited tumor cell migration and invasion compared with controls (p<0.0001).

Fig 5: HMGA1 promotes HBV gene expression and replication in HBV-infected HepG2/NTCP cells. HepG2/NTCP cells were infected with HBV virions at a MOI of 1000 geq/cell overnight. Culture media were changed the next day, and at day 2, cells were transduced by shHMGA1-expressing lentivirus or control virus (Lenti-shHMAG1 or Lenti-shCtrl) (A and B), or Lenti-HMGA1-Flag or control virus Lenti-Ctrl (A and C). Culture media were changed at day 3, and then every 2 days. At day 7, intracellular HBV RNA, core-associated DNA and cccDNA were examined using northern and southern blot, and qrtPCR, respectively. Endogenous and exogenous HMGA1 were determined in western blot using anti-HMGA1 and anti-Flag antibodies, respectively. HBsAg and HBeAg in culture media were assayed using ELISA. The relative levels of HBV RNA and DNA were quantified using densitometry scanning. Group means and SEMs of normalized values within transfection group were presented. Data of Lenti-shHMGA1or Lenti-HMGA1-Flag-transduced groups were compared against control virus group using unpaired two-tailed t-test; *, P < 0.05; **, P < 0.01; ***, P < 0.001.