Extended Data Fig. 6: Characterization of Haster mutants.
From: The HASTER lncRNA promoter is a cis-acting transcriptional stabilizer of HNF1A
a, Body weight of males at 8 weeks of age. HasterpKO (n = 8), Pdx1-Cre;Haster+/+ (n = 12) and Hasterf/f (n = 8). Mean ± s.d. b, Intraperitoneal glucose tolerance test in 8-week-old and 30-week-old female HasterpKO (n = 9), Pdx1-Cre;Haster+/+ (n = 10) and Hasterf/f (n = 10). Mean ± s.e.m., *P ≤ 0.05, **P ≤ 0.01 and ***P ≤ 0.001 (two-tailed Student’s t-test). c, Body weight of 8- to 10-week-old male Haster-/- (n = 9), Haster+/- (n = 12) and Haster-/- (n = 13). Mean ± s.d. d, Intraperitoneal glucose tolerance test in 8- to 10-week-old females Haster-/- (n = 10), Haster+/- (n = 10) and Haster+/+ (n = 12). Mean ± s.e.m., *P ≤ 0.05, **P ≤ 0.01 and ***P ≤ 0.001 (two-tailed Student’s t-test). e, Relative quantification of HNF1A-negative β cells at the indicated age and genotype. Results show that HNF1A silencing correlates with time of Haster knockout, with higher silencing frequency after early deletion (Haster germline KO and HasterpKO models). HNF1A silencing increased with time in β cells from germline KO and HasterpKO models, but not when excision occurred in early β cells (βKO). No HNF1A silenced β cells were observed after Pdx1-CreERTM-based tamoxifen-inducible excision in adult β cells (HasteriβKO model). Mean ± s.d from 2-4 sections per mouse, 2-4 mice per condition. f-h, Immunofluorescence for HNF1A and insulin in E15.5 (f) Haster+/+, (g) HasterpKO, and (h) Haster-/- pancreas. Solid arrowheads, insulin cells overexpressing HNF1A; hollow arrowheads, insulin cells lacking HNF1A. i-k, Immunofluorescence for HNF1A and insulin in adult (i) wild-type, (j) HasterβKO and (k) HasteriβKO pancreas. Arrows point to HNF1A-negative β cells. Most β cells from HasterβKO and HasteriβKO islets overexpress HNF1A. e-k, n = 2 wild type embryos and n = 3 adult wild type mice; n = 2 Haster-/- embryos and adult mice; n = 3 HasterpKO embryos and adult mice; n = 2 HasterβKO 6- and 32-week-old mice; n = 2 HasteriβKO mice. n.a.: not analyzed because deletions were performed at a later time point. Scale bar, 50 µM.
