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. 2020 May 13;12(5):1227. doi: 10.3390/cancers12051227

Figure 3.

Figure 3

LINC00261 regulation by its genomic neighbor FOXA2. (a) The genomic loci of LINC00261 and FOXA2 on chromosome 20. (b) The siRNA-mediated knockdown of FOXA2 significantly downregulated FOXA2 and LINC00261 RNA levels (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, one-way ANOVA). (c) Stable overexpression of FOXA2 in PANC-1 was achieved at RNA (left panel; glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as the reference gene) and protein levels (right panel; ribosomal protein L7 (RPL7) was used as a loading control; **** p < 0.0001, unpaired t-test). (d) LINC00261 expression levels in control or FOXA2 overexpressing PANC-1 cells (* p < 0.05, unpaired t-test). (e) Luciferase activity of a LINC00261 promoter reporter after stable FOXA2 or empty control vector overexpression in PANC-1 cells (**** p < 0.0001, unpaired t-test). (f) Chromatin immunoprecipitation (ChIP) followed by qPCR analysis using primers located upstream (1,2) and downstream (3) of the LINC00261 transcriptional start site (upper panel) confirmed binding of FOXA2 to the LINC00261 promoter region (lower panel, **** p < 0.0001, unpaired t-test).