Fig. 5: The HASTER promoter is a positive and negative cis-acting element.
From: The HASTER lncRNA promoter is a cis-acting transcriptional stabilizer of HNF1A
a, Severe fasting and fed hyperglycaemia (left; n = 12 wild-type (WT) mice, n = 10 Haster+/− mice, n = 11 Hnf1a+/− mice and n = 13 Hnf1a+/−;Haster+/− mice) and reduced insulin secretion (right; n = 5 mice per genotype) in Hnf1a+/−;Haster+/− compound heterozygotes. The data are presented as means ± s.d. Statistical significance was determined by two-tailed Student’s t-test. b, Immunofluorescence showing normal HNF1A in Hnf1a+/− islets and no expression in most islet cells from adult Hnf1a+/−;Haster+/− mice (n = 1 per genotype). Solid arrowhead: HNF1Ahigh acinar cell. Hollow arrowhead: HNF1Alow β cell. Scale bar, 50 µm. c, Allele-specific Hnf1a mRNA in islets from hybrid-strain mice carrying the Haster mutation in the C57BL/6 chromosome. Hnf1a was quantified by strain-specific qPCR and normalized to Tbp (n = 4 mice per genotype). The data are presented as means ± s.d. Statistical significance was determined by two-tailed Student‘s t-test, d, Strain-specific RNA-seq analysis from Haster+/stop and Haster+/+ PWK/PhJ;C57BL/6 hybrid islets (n = 4 mice per genotype). RPM, reads per million reads. e, HNF1A overexpression in liver from Hnf1a+/−;Haster+/− mice (n = 1 per genotype). Scale bar, 50 µm. f, Allele-specific Hnf1a mRNA in liver from Haster+/− hybrid-strain mice carrying the Haster mutation in the C57BL/6 chromosome. Hnf1a was quantified with strain-specific assays and normalized to Tbp (n = 4 mice per genotype). The data are presented as means ± s.d. Statistical significance was determined by two-tailed Student’s t-test. g, Strain-specific RNA expression from Haster+/stop C57BL/6;PWK/PhJ hybrid mice, showing that reducing Haster elongation in liver failed to increase Hnf1a expression from the same C57BL/6 allele. The graphs show reads per million (RPM) (means ± s.d.). h, Targeting dCAS9 to the HASTER transcriptional start site blocked HASTER transcription in EndoC-βH3 cells but did not affect HNF1A or HNF4A mRNAs (n = 3 lentiviral transductions). i, CRISPR–SAM HASTER activation in EndoC-βH3 cells did not affect HNF1A and HNF4A (n = 3 lentiviral transductions). In h and i, the data represent normalized expression levels (means ± s.d.) and statistical significance was determined by two-tailed Student’s t-test.
