Fig. 4: Haster deletion causes islet cell HNF1A hyperactivation or silencing and diabetes.
From: The HASTER lncRNA promoter is a cis-acting transcriptional stabilizer of HNF1A
a, Intraperitoneal glucose tolerance in 8-week-old male mice (n = 8 HasterpKO, n = 12 Pdx1-Cre;Haster+/+ and n = 8 Hasterf/f). P = 0.045, 8 × 10−3, 3 × 10−4, 4 × 10−4 and 5 × 10−3 at 0, 15, 30, 60 and 120 min, respectively. b, Plasma insulin of 8-week-old male mice (n = 7 HasterpKO and n = 6 Pdx1-Cre;Haster+/+). P = 0.83, 2 × 10−3 and 3 × 10−4 at 0, 15 and 30 min, respectively. c, Intraperitoneal glucose tolerance in 8-week-old male mice (n = 9 Haster−/−, n = 12 Haster+/− and n = 13 Haster+/+). P = 0.048, 0.075, 0.011, 4 × 10−4 and 2 × 10−4 at 0, 15, 30, 60 and 120 min, respectively. d, Plasma insulin in 8-week-old male mice (n = 7 Haster−/−, n = 6 Haster+/+ and n = 6 Haster+/−). P = 0.042, 0.045 and 0.026 at 0, 15 and 30 min, respectively. e, Glucose-to-insulin ratio in 8-week-old male mice (n = 9 Haster−/−, n = 12 Haster+/− and n = 13 Haster+/+). In a–e, the data are presented as means ± s.e.m. and statistical significance was determined by two-tailed Student’s t-test (*P ≤ 0.05; **P ≤ 0.01; ***P ≤ 0.001). f, Immunofluorescence for HNF1A and insulin, showing either HNF1A overexpression (solid arrowheads) or no HNF1A expression (empty arrowheads) in endocrine cells of adult HasterpKO and Haster−/− mice. Note that all acinar cells from mutant mice overexpressed HNF1A (n = 3 Haster+/+, n = 3 HasterpKO and n = 2 Haster−/−). g, Immunofluorescence for HNF1A, PDX1 (a pancreatic and duodenal marker) and glucagon in Haster−/− and control E11.5 embryos, showing low heterogeneous HNF1A in pancreatic but not gut progenitors. dp, dorsal pancreas (delineated by dashed lines in KO); du, duodenum. h, Kernel density estimation of HNF1A-regulated gene expression (average z score) showing either down- or upregulation of HNF1A-dependent genes in HasterpKO HNF1Alow and HNF1Ahigh β cell clusters. i, RNA-seq (RPKM) from the indicated hESC-derived differentiation stages. j, HNF1A mRNA in hESC-derived pancreatic progenitors carrying HASTER P1 homozygous deletions (see Fig. 2a) (n = 5 independent differentiations). The data are presented as TBP-normalized relative expression (means ± s.d.). Statistical significance was determined by two-tailed Student’s t-test. k, Immunofluorescence for HNF1A, PDX1 and NKX6-1 in hESC-derived pancreatic progenitors carrying the indicated deletions, showing downregulation of HNF1A (n = 2 per deletion). In f, g and k, the scale bars represent 50 µm.
