Fig 2: Effects of Wnt3a silencing on Eaf2 expression in HLE-B3 cells. (A) Representative images of Annexin V/PI uptake in Eaf2-overexpressing HLE-B3 cells following Wnt3a knockdown. (B) Quantification of apoptotic cells in Eaf2-overexpressing HLE-B3 cells following Wnt3a knockdown. (C) Quantification of LDH activity in Eaf2-overexpressing HLE-B3 cells following Wnt3a knockdown. Reverse transcription-quantitative polymerase chain reaction was conducted to determine the effects of Wnt3a silencing on the mRNA expression levels of (D) caspase 3, Eaf2 and Wnt3a, and (E) β-catenin, Bcl-2 and Bax in HLE-B3 cells overexpressing Eaf2. Western blot analysis was conducted to determine the effects of Wnt3a silencing on the protein expression levels of (F) β-catenin, p-GSK3β, GSK3β, Bcl-2 and Bax, and (G) caspase 3 and Wnt3a in HLE-B3 cells overexpressing Eaf2. *P<0.05, **P<0.01 vs. control. Group 1 represents the oeEaf2 group consisting of HLE-B3 cells overexpressing Eaf2; group 2 represents the oeEaf2 + shWnt3a group consisting of Eaf2-overexpressing cells transfected with Wnt3a shRNA; group 3 represents the oeEaf2 + shCon group consisting of Eaf2-overexpressing cells transfected with control shRNA. 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; HLE, human lens epithelial; LDH, lactate dehydrogenase; p, phosphorylated; PI, propidium iodide; shRNA, short hairpin RNA.

Fig 3: Overview of the involvement of LINC00301 in NSCLC. Schematic representation of our proposed model of FOXC1 upregulates LINC00301 expression, the latter partially mediates oncogenic effects through its epigenetic silencing of EAF2 expression by directly binding with EZH2 to mediate HIF1a activation in the nucleus and by acting as ceRNA for miR-1276 to increase HIF1a expression in the cytoplasm in NSCLC

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: LINC00301 facilitates proliferation, migration, and invasion through regulation of EAF2/VHL/HIF1a pathway in NSCLC cells. a Represent images for EAF2, pVHL, HIF1a in LINC00301 KD, and LINC00301 OE A549 and SPC-A-1 cells analyzed by immunofluorescence. b IB tested EAF2, pVHL, HIF1a levels in sh-NC, sh-1, sh-EAF2, sh-1 + sh-EAF2 transfected A549, and SPC-A-1 cells. c IB tested EAF2, pVHL, HIF1a levels in pLenti-CMV-vector, pLenti-CMV-LINC00301, pLenti-CMV-EAF2 transfected in A549, and SPC-A-1 cells. d Colony formation assays (seeded in 6-well plate) tested cell proliferation in sh-NC, sh-1, sh-EAF2, sh-1 + sh-EAF2, pLenti-CMV-vector, pLenti-CMV-LINC00301, pLenti-CMV-EAF2 or pLenti-CMV-LINC00301 + pLenti-CMV-EAF2 transfected in A549 and SPC-A-1 cells. e Trypan blue staining tested cell viability in sh-NC, sh-1, sh-EAF2, sh-1 + sh-EAF2, pLenti-CMV-vector, pLenti-CMV-LINC00301, pLenti-CMV-EAF2 or pLenti-CMV-LINC00301 + pLenti-CMV-EAF2 transfected in A549 and SPC-A-1 cells. g Transwell migration assay tested migration activity in sh-NC, sh-1, sh-EAF2, sh-1 + sh-EAF2, pLenti-CMV-vector, pLenti-CMV-LINC00301, pLenti-CMV-EAF2 or pLenti-CMV-LINC00301 + pLenti-CMV-EAF2 transfected in A549 and SPC-A-1 cells. f Transwell migration assay tested migration activity in sh-NC, sh-1, sh-EAF2, sh-1 + sh-EAF2, pLenti-CMV-vector, pLenti-CMV-LINC00301, pLenti-CMV-EAF2 or pLenti-CMV-LINC00301 + pLenti-CMV-EAF2 transfected in A549 and SPC-A-1 cells. g Transwell invasion assay tested invasive activity in sh-NC, sh-1, sh-EAF2, sh-1 + sh-EAF2, pLenti-CMV-vector, pLenti-CMV-LINC00301, pLenti-CMV-EAF2 or pLenti-CMV-LINC00301 + pLenti-CMV-EAF2 transfected in A549 and SPC-A-1 cells. Assays were conducted in triplicate. *p < 0.05, means ± SD was shown. Statistical analysis was performed by Student’s t-test