Fig 2: The schematic diagram of this study. MNX1-AS1 promotes LSCC cell growth and migration through targeting miR-744-5p to activate the BCL9/β-catenin signaling. LSCC: laryngeal squamous cell carcinoma.

Fig 3: MNX1-AS1 increased BCL9 expression via targeting miR-744-5p. (A) The putative binding sites between miR-744-5p and BCL9. (B) The luciferase activity of BCL9-wt/mut was detected by luciferase gene reporter Assay (n = 3, **P < 0.01, compared with NC mimic group). (C) BCL9 protein levels in NC mimic or miR-744-5p mimic transfected cells were detected by using Western blotting (n = 3, **P < 0.01, compared with NC mimic group). (D) BCL9 protein levels in AMC-HN-8 and TU686 cells in control+NC mimic, MNX1-AS1+NC mimic and MNX1-AS1+miR-744-5p mimic groups were detected by Western blotting (n = 3, ** P < 0.01, compared with control+NC mimic group; ## P < 0.01, compared with mnx1-as1+nc mimic group). (E) QRT-PCR was used to detect the mRNA level of BCL9 in human LSCC tissues and the paired normal tissues (n = 65, ** P < 0.01, compared with normal group). (F) Correlations between the expression levels of miR-744-5p and BCL9/MNX1-AS1 in 65 cases of human LSCC tissues were determined by Pearson Correlation Analysis.

Fig 4: Mouse dental pulp tissues cultured under hypoxic culture show increased expression of Hif1α and Bcl9, and nuclear co-localization of Bcl9 and β-catenin. (a) Immunofluorescence detection of Hif1α in mouse dental pulp tissue in hypoxic culture (1% O2) for 48 h. Hif1α expression is increased and detected in the nucleus. (b) Immunofluorescence detection of Bcl9 and β-catenin in mouse dental pulp tissue in hypoxic culture (1% O2) for 48 h. Double immunofluorescence staining for BCL9 and β-catenin shows co-localization.

Fig 5: BCL9 overexpression promotes osteo/odontoblast differentiation and Wnt/β-catenin signaling. (a–c) BCL9 overexpression upregulates the mRNA expression of BCL9, Wnt/β-catenin target genes (AXIN2 and TCF1), and osteo/odontoblast differentiation markers (RUNX2, ALP, and OC) for 24 h. (d) TCF/LEF luciferase assay showing the activation of Wnt/β-catenin signaling in hDPSCs with BCL9 overexpression for 24 h. (e) Immunoprecipitation to detect the binding of BCL9 to β-catenin in BCL9 (FLAG Tag) and β-catenin (V5 Tag) overexpression for 24 h. (f) BCL9 and β-catenin protein expression are upregulated in the nucleus of hDPSCs in BCL9 and β-catenin overexpression for 24 h. Full-length blots are presented in Supplementary Fig. 2. Error bars indicate standard deviation (n = 4). *p < 0.05.