Fig. 2: HASTER negative feedback regulates HNF1A in mice and humans.
From: The HASTER lncRNA promoter is a cis-acting transcriptional stabilizer of HNF1A
a, Homozygous deletions of the HASTER promoter (two deletions with independent sgRNA pairs) or control deletions in HNF1A intron 1 or AAVS1 were generated in hESCs. b, HNF1A mRNA was increased in differentiated hepatocytes from HASTER mutant hESCs (n = 3 independent clones per deletion). The bar graphs show RPLP0-normalized expression values (means ± s.d.). Statistical significance was determined by two-tailed Student’s t-test. Act. A, Activin A; BMP-4, bone morphogenetic protein 4; HGF, hepatocyte growth factor; OSM, Oncostatin M. c, Schematic of the mouse Hasterf allele. d, Liver RNA levels in seven HasterLKO and eight control mice. The data represent Tbp-normalized values (means ± s.d.). Statistical significance was determined by two-tailed Student’s t-test. e, Liver HNF1A immunofluorescence in the indicated genotypes. Scale bar, 50 µm. f, Western blot for HNF1A on liver extracts (n = 3 mice for each genotype). The bars represent relative expression levels (means ± s.d.). Statistical significance was determined by two-tailed Student’s t-test. Ctrl, control. g,h, Haster was decreased in Hnf1a−/− islets (g; n = 4 Hnf1a−/− and n = 5 Hnf1a+/+ mice) and liver (h; n = 4 mice per genotype). The bars represent relative expression levels (means ± s.d.). Statistical significance was determined by two-sided Wald test with adjusted P values. i, EndoC-βH3 cells carrying an indel in HNF1A exon 1 showed decreased HASTER as well as HNF4A—another HNF1A-dependent gene (n = 3 lentiviral transductions). The data represent means ± s.d. and are normalized to TBP. Statistical significance was determined by two-tailed Student’s t-test. j, HNF1A binds the Haster promoter in mouse liver (representative example from three replicates, MACS2 P values). The locations of seven HNF1A motifs with a JASPAR CORE score of >0.8 are shown, along with the sequences of three motifs. See Extended Data Fig. 1 for information on transcriptional start sites. k, Schematic of the HNF1A/HASTER negative feedback loop.
