Fig 2: SNHG12 promotes GC proliferation in vivo. (A, B) Comparison of tumor formation between the NC groups and sh-SNHG12 groups. (C) SNHG12 and Ki-67 levels between the NC groups and sh-SNHG12 groups by ISH and IHC. (D–H) Expression of YWHAZ, AKT, p-AKT, GSK-3β, and p-GSK-3β proteins in the NC and sh-SNHG12 groups by IHC. (I) Schematic illustration of the mechanism underlying SNHG12 regulation of GC proliferation. Significant results were presented as ***P < 0.001. Magnification ×200, magnification ×400, scale bar 20,000 nm.

Fig 3: SNHG12 acts as a ceRNA for miR-218-5p to regulate YWHAZ expression in GC cells. (A, B) FISH assays and cytoplasmic and nuclear RNA purification assays indicate that SNHG12 is located in GC cell cytoplasm. (C, D, E) qRT-PCR assays showing relative expression of miR-218-5p, SNHG12 and YWHAZ in GC cells transfected with miR-218-5p mimics or inhibitors. (F, G) Luciferase assays revealed the interaction between miR-218-5p and SNHG12. (H, I) Luciferase assays revealed the interaction between miR-218-5p and YWHAZ. (J, K, L, M, N, O) qRT-PCR assays indicated the relative expression of SNHG12 and YWHAZ in GC cells transfected with miR-218-5p mimics or pCDH-CMV-SNHG12 and sh-SNHG12 or si-YWHAZ or miR-218-5p inhibitors. (P, Q, R) RIP assays indicated the binding of SNHG12, miR-218-5p, or YWHAZ with Ago2. Significant results were presented as *P<0.05, **P<0.01, ***P<0.001. Magnification ×200, Scale bar 50 µm. Magnification ×400, Scale bar 20 µm.

Fig 4: SNHG12 increases the expression of ß-catenin via YWHAZ stabilizing ß-catenin and stabilizing CTNNB1 mRNA. (A, B) The RNA and protein expressions of ß-catenin after YWHAZ knockdown were verified by qRT-PCR and WB. (C, D) The RNA and protein expressions of ß-catenin after SNHG12 knockdown were verified by qRT-PCR and WB. (E) Co-IP assays showing the interaction between YWHAZ and ß-catenin. (F, G) Ubiquitination changes after knockdown YWHAZ and SNHG12 tested by IP in GC cells. (H) RIP assays showing SNHG12 binding with HuR. (I) Interaction probabilities between HuR and CTNNB1 predicted by RPIseq online tool. (J) RIP assays showing CTNNB1 binding with HuR. (K, L) The stability of CTNNB1 after knockdown SNHG12 or HuR. (M, N) The nuclear expression of ß-catenin after SNHG12 knockdown and overexpression was tested by western blotting. (O) Luciferase assays showing the effects on TOP/FOP reporter activity in MGC-803 and AGS cells with SNHG12 overexpression. Significant results were presented as *P<0.05, **P<0.01, ***P<0.001. No significantly differences were presented as ns.

Fig 5: Association of mRNA expression of 14-3-3 isoforms with clinical stages of BrCa. (A) Summarization of transcriptional expression of 14-3-3 isoforms in different clinical stages. (B, E, G) Expression levels of 14-3-3 beta, 14-3-3 zeta and 14-3-3 theta were related to clinical stages. (C, D, F, H) Expression levels of 14-3-3 gamma, 14-3-3 epsilon, 14-3-3 eta and 14-3-3 sigma were not associated with clinical stages.