Supplementary Figure 1 - Ling
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1 HA1 (%) Insulin seretion (SI) e 7. p= p= a-ell granules -ell granules BMI (kg/m ) -ells / (a- + -ells) p= p= Age: -ell ontent: % Age: -ell ontent: % Age: -ell ontent: % Age:7 -ell ontent : % Supplementary Figure 1. Age orrelates positively with HA1 levels. Correlation etween age an HA1 levels (a), BMI (), insulin seretion (), an islet -ell ontent () in panreati islet onors as analyse y linear regression analysis (a-, n=7) or Spearman orrelation (, n=1). (e) Representative eletron mirographs of panreati islets from four onors, ages an a-ell granules are iniate y re an yellow arrowheas, respetively. Sale ar in the lower right panel is mm. Supplementary Figure 1 - Ling
2 g17 g Islets GNPNAT1 q=.x1-7 Islets q=1.x1-1 7 g17 g 1 Bloo 7 GNPNAT1 p=1.x Bloo p=. 1 7 g17 g Bloo p=. 7 Bloo GNPNAT1 p=.7 g1 Islets q=.x1-1 7 g1 1 Bloo p=. 1 7 g1 Bloo 1 p=. 7 Supplementary Figure. Age-assoiate DNA methylation hanges of GNPNAT1 an in human panreati islets an loo. The assoiation etween age an DNA methylation of sites in GNPNAT1 (a) an () in panreati islets from the human islet ohort (n=7, left panels), an loo from the Danish Family Stuy (n=11) at aseline (mile panels) an follow-up (right panels). Data were analyse y linear regression. Supplementary Figure - Ling
3 g717 g g71 1 KLF1 p=.x1-7 7 FHL p=.x FAM1C p=.x1-1 7 g7 g7 g17 1 KLF1 p=. 7 7 FHL p=.x1-7 ZNF1B 1 p=.1x1-7 g77 g1 1 1 FHL p=7.x1-7 ZNF1B p=1.7x1-7 Supplementary Figure. DNA methylation of CpG sites in KLF1, FHL, FAM1C, an ZNF1B at follow-up in loo from the Danish Family Stuy. Pyrosequening of DNA from onors in the Danish Family Stuy reveale that methylation of CpG sites in KLF1 (a), FHL (), FAM1C (), an ZNF1B () assoiate signifiantly with age also at follow-up as analyse y linear regression (n=11). Supplementary Figure - Ling
4 KLF1 1 g717 g7 FHL g g7 g77 FAM 1C 1 g ZNF1B 1 g17 g1 Supplementary Figure. DNA methylation of KLF1, FHL, FAM1C, an ZNF1B in loo inrease over time. The ifferene in methylation of iniate sites in loo samples from the Danish Family Stuy (n=11) at aseline an at follow-up was analyse with a Wiloxon signe rank test. p <.1. Supplementary Figure - Ling
5 1 g717 KLF1 Bloo Panreas g7 p=.x1-11 g117 g FHL g7 g77 FAM1C g71 g17 ZNF1B g1 7 Bloo 1 1 Islets g717 g7 g117 g g7 g77 g71 g17 g1 Supplementary Figure. DNA methylation of CpG sites in KLF1, FHL, FAM1C, an ZNF1B are similar in panreas an loo. (a) Analysis of pulishe ata y Slieker et al show that methylation of the stuie sites in KLF1, FHL, FAM1C, an ZNF1B is similar in panreas an loo taken from the same onors (n=). GNPNAT1 an were exlue from this analysis as they i not exhiit similar age-assoiate hanges in DNA methylation in our islet an loo ata. () Methylation of the nine sites in KLF1, FHL, FAM1C, an ZNF1B in panreas an loo taken form the same onors orrelate signifiantly. Supplementary Figure - Ling
6 CpG e 1 KLF1 p=.x FHL p=1.x FHL p=.x FAM1C p=.x ZNF1B p=.x GNPNAT1 p=7.x1-1 7 CpG CpG CpG CpG CpG f 1 KLF1 p=1.x FHL p=.x FHL p=1.1x FAM1C p=1.x1-1 7 ZNF1B 1 p=.x p= CpG CpG CpG CpG KLF1 p=.x FHL p=1.1x FAM1C p=1.x CpG ZNF1B p=.x1-1 7 p=. 1 7 Supplementary Figure. DNA methylation of surrouning CpG sites in KLF1, FHL, FAM1C, ZNF1B an GNPNAT1 assoiates with age at aseline in the Danish Family Stuy. Pyrosequening reveale that several surrouning sites in KLF1 (a), FHL (), FAM1C (), ZNF1B (), an GNPNAT1 (e) were signifiantly assoiate with age in loo at aseline (n=11). However, the surrouning sites in (f) i not assoiate with age at aseline. Data were analyse y linear regression. Supplementary Figure - Ling
7 e CpG KLF1 p=.7x FHL p=1.x FHL p=1.x1-7 FAM1C p=.x1-1 7 ZNF1B p= GNPNAT1 p=.x1-7 CpG CpG CpG CpG CpG f KLF1 1 p=1.x1-7 7 FHL p=.x1-7 1 FHL p=.x1-7 FAM1C 1 p=1.x1-7 ZNF1B p= p=. 7 CpG CpG CpG CpG CpG KLF1 p=.x FHL p=.x1-7 FAM1C p=.x1-1 7 ZNF1B p= p=. 7 Supplementary Figure 7. DNA methylation of surrouning CpG sites in KLF1, FHL, FAM1C, an GNPNAT1 assoiates with age at follow-up in the Danish Family Stuy. Pyrosequening reveale that several surrouning sites in KLF1 (a), FHL () FAM1C (), an GNPNAT1 (e) were signifiantly assoiate with age in loo at follow-up (n=11). However, the surrouning sites in ZNF1B () an (f) i not assoiate with age at follow-up. Data were analyse y linear regression. Supplementary Figure 7 - Ling
8 g717 KLF1 1 CD1+ Monoytes CD+ NK ells CD+ T ells g7 KLF1 1 1 CD1+ Monoytes CD+ NK ells CD+ T ells g FHL 7 1 CD1+ Monoytes CD+ NK ells CD+ T ells g7 1 FHL CD1+ Monoytes CD+ NK ells CD+ T ells g77 FHL 7 1 CD1+ Monoytes CD+ NK ells CD+ T ells g71 FAM1C 1 CD1+ Monoytes CD+ NK ells CD+ T ells g17 ZNF1B 1 1 CD1+ Monoytes CD+ NK ells CD+ T ells g1 ZNF1B 1 CD1+ Monoytes CD+ NK ells CD+ T ells g17 1 GNPNAT1 CD1+ Monoytes CD+ NK ells CD+ T ells Supplementary Figure. DNA methylation of KLF1, FHL, FAM1C, ZNF1B, an GNPNAT1 in isolate human loo ell populations. Pulishe methylation ata y Reinius et al from isolate loo ell populations suggest that the age-assoiate methylation hanges ientifie in KLF1, FHL, FAM1C, ZNF1B, an GNPNAT1 are not ue to an altere loo ell omposition (n=). Supplementary Figure - Ling
9 g117 g g71 KLF1 p=.x FHL p=.x FAM1C 1 p=1.x1-1 7 g717 g7 KLF1 p=.x FHL p=.x g117 < years > years KLF1 g717 * g7 g FHL g7 g7 g77 KLF1 1 p=.x1-1 7 FHL p=1.x g77 g71 FAM1C ** Supplementary Figure. Biologial repliation of age-assoiate DNA methylation hanges in human panreati islets. Pyrosequening analysis of methylation of seven age-assoiate CpG sites in KLF1 (a), FHL (), an FAM1C () on DNA from panreati islets from a repliation ohort of onors verifie that methylation of all analyse CpG sites orrelate signifiantly with age. Data were analyse with Spearman orrelations. () DNA methylation iffere signifiantly for all seven sites in panreati islets from young (< years, n=) versus ol (> years, n=1) onors in the valiation ohort. Data analyse with a Mann-Whitney U test. *p<., **p<.1, an p<.1. Supplementary Figure - Ling
10 1 Purity of islet preparations (%) Supplementary Figure 1. There was no impat of age of the onor on the purity of islet preparations, thus exluing suh a ias as onfouning fator. Supplementary Figure 1 - Ling
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