Figure 7

LncRNA TINCR mediates the function of ZNF750. (a) qRT-PCR analysis of TINCR in SCC cells either with ectopic expression of GFP control (CTL) or wildtype ZNF750 and (b) in ME180 cells (CSCC) transfected with either scrambled siRNAs (siCTL) or ZNF750 siRNAs (siZNF750). Data show mean±s.d. N=3–6. *P<0.05. (c) Co-expression analysis of TINCR transcripts and ZNF750 protein (40 μg/lane) levels in variety of SCC cell lines. (d) Human H1 embryonic stem cells were differentiated to keratinocyte progenitors in vitro, and the global RNA expression profile during differentiation were analysed using RNA-seq. Sequencing reads after normalization spanning ZNF750 gene are shown. Numbers indicate FPKM value (Fragments Per Kilobase of Exon per Million Fragments Mapped). (e) Co-expression analysis of ZNF750 mRNA and TINCR transcripts from TCGA dataset. (f) Summary of correlation coefficient between ZNF750 mRNA and TINCR transcripts across different tumour types from TCGA dataset. (g) SCC cells expressing either GFP (CTL, control) or wildtype ZNF750 were transfected with either scrambled or TINCR siRNAs and then subjected to foci formation assay (representative photos), and (h) foci were quantified. Data show mean±s.d. N=4. *P<0.05. (i) SCC cells with indicated treatment were subjected to migration assay. (j) Effects of TINCR knockdown on differentiation-related gene expression; heatmaps show average fold change measured by qRT-PCR (N=3). (k) Low TINCR expression is associated with poor disease-free survival of HNSCC patients on TCGA cohort.