Fig 2: H2O2-treated HLE-B3 cells overexpressing Eaf2, as observed by fluorescence microscopy. Immunocytochemistry was used to examine Wnt3a expression in H2O2-treated Eaf2-overexpressing HLE-B3 cells following Wnt3a knockdown. The oeEaf2 group consists of HLE-B3 cells overexpressing Eaf2; the oeEaf2 + shWnt3a group consists of Eaf2-overexpressing cells transfected with Wnt3a shRNA; the oeEaf2 + shCon group consists of Eaf2-overexpressing cells transfected with control shRNA. Eaf2, ELL-associated factor 2; H2O2, hydrogen peroxide; HLE, human lens epithelial; shRNA, short hairpin RNA.

Fig 3: HLE-B3 cells as observed by fluorescence microscopy. Immunocytochemistry techniques were used to examine the expression levels of (A) Eaf2 and (B) Wnt3a in hydrogen peroxide-treated HLE-B3 cells overexpressing Eaf2. Con represents the control group consisting of normal HLE-B3 cells; oeCon represents the negative control group consisting of HLE-B3 cells transfected with empty pcDNA3.0 plasmid; oeEaf2 represents the experimental group consisting of HLE-B3 cells transfected with pcDNA3.0-Eaf2. Eaf2, ELL-associated factor 2; HLE, human lens epithelial.

Fig 4: Knockdown of SNHG22 inhibits GC progression in vivo.A Subcutaneous tumors were separated and imaged at the endpoint of the experiment (n = 6). B Tumor growth curves of different subcutaneous tumor groups are shown. C Relative expression of SNHG22 in subcutaneous tumor tissues were quantified using qRT-PCR. D Relative expression of indicated genes in subcutaneous tumor tissues were quantified using western blot. E The expression levels of ki-67, Notch1 and E-cadherin in different groups of subcutaneous tumors were evaluated using IHC. F–K The inverse correlation between SNHG22 and Notch1, E-cadherin, EAF2, ADRB2, rap1GAP, RUNX3 expression in human GC tissues was analyzed using IHC.

Fig 5: Effects of Eaf2 overexpression on H2O2-induced apoptosis of HLE-B3 cells. (A) Representative images of Annexin V/PI uptake by H2O2-induced HLE-B3 cells following Eaf2 overexpression, as determined by flow cytometry. (B) Quantification of apoptotic cells in H2O2-induced HLE-B3 cells overexpressing Eaf2. (C) Quantification of LDH activity in H2O2-induced HLE-B3 cells overexpressing Eaf2. Reverse transcription-quantitative polymerase chain reaction indicating the effects of Eaf2 overexpression on the mRNA expression levels of (D) caspase 3, Wnt3a and Eaf2, and (E) β-catenin, Bcl-2 and Bax in H2O2-treated HLE-B3 cells. Western blot analysis indicating the effects of Eaf2 overexpression on the protein expression levels of (F) β-catenin, p-GSK3β, GSK3β, Bcl-2 and Bax, and (G) Wnt3a, Eaf2 and caspase 3 in H2O2-treated HLE-B3 cells. *P<0.05, **P<0.01, ***P<0.001 vs. Con or oeCon. Con represents the control group consisting of normal HLE-B3 cells; oeCon represents the negative control group consisting of HLE-B3 cells transfected with empty pcDNA3.0 plasmid; oeEaf2 represents the experimental group consisting of HLE-B3 cells transfected with pcDNA3.0-Eaf2. Bax, B-cell lymphoma 2-associated X protein; Bcl-2, B-cell lymphoma 2; GSK3β, glycogen synthase kinase 3β; Eaf2, ELL-associated factor 2; FITC, fluorescein isothiocyanate; H2O2, hydrogen peroxide; HLE, human lens epithelial; LDH, lactate dehydrogenase; p, phosphorylated; PI, propidium iodide.