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Sandra Hellberg, MSc, Ratnesh B

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1 Maintained thymic output of conventional and regulatory T cells during human pregnancy 
Sandra Hellberg, MSc, Ratnesh B. Mehta, PhD, Anna Forsberg, PhD, Göran Berg, MD, PhD, Jan Brynhildsen, MD, PhD, Ola Winqvist, MD, PhD, Maria C. Jenmalm, PhD, Jan Ernerudh, MD, PhD  Journal of Allergy and Clinical Immunology  Volume 143, Issue 2, Pages e7 (February 2019) DOI: /j.jaci Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 TREC levels in pregnant and nonpregnant women. A, Representative flow cytometry dot plot for sorting strategy. B, Foxp3 mRNA expression determined by RT-PCR in sorted T-cell subpopulations (n = 4 nonpregnant, n = 4 pregnant). TREC levels in sorted (C) memory Treg and Tconv cells compared with naive T-cell subsets, (D) naive Treg cells, and (E) naive Tconv cells, determined by RT-PCR using GAPDH as a reference gene. Missing values are due to limited availability in the number of cells required for analysis. GAPDH, Glyceraldehyde-3-phosphate dehydrogenase; n, naive; m, memory. Median and interquartile ranges are shown. *P < .05 and ****P < .0001. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 RTE (CD31+CD45RA+) subsets in pregnant and nonpregnant women. A, Representative flow cytometry dot plots for gating strategies. Proportion and absolute numbers of RTE Tconv cells (B and C) and RTE Treg cells (D and E) analyzed by flow cytometry. Missing values are due to limited availability in the number of cells required for analysis. Mean and SDs are shown. **P < .01. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig E1 Gating strategy phenotyping by flow cytometry. A, Subpopulations of CD4+ cells were defined by CD45RA and Foxp3 expression and further analyzed for expression of (B) CD127, (C) CD31, and (D) Ki67. Representative flow cytometry dot plots from 1 individual are shown. MN, Mature naive. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig E2 Correlation between TREC levels and CD31 expression. TREC levels and CD31 expression in (A) naive and memory Tconv cells (n = 9) and (B) naive and memory Treg cells (n = 9). TREC levels were determined by RT-PCR, and CD31 expression was analyzed by flow cytometry. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig E3 Ki67 expression on CD4+ T-cell subsets. Proportion of Ki67+ (A) mature naive and memory Tconv and Treg cells and nonsuppressive Foxp3+ TH cells, (B) RTE T conv cells, and (C) RTE Treg cells, analyzed by flow cytometry. Mean and SDs are shown. Missing values are due to limited availability in cell numbers required to perform analysis. *P < .05, ***P < .001, ****P < .0001. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig E4 Proportion of T-cell subsets. Frequency of (A and B) mature naive Tconv and Treg cells, (C and D) memory Tconv and Treg cells, and (E) nonsuppressive Foxp3+ TH cells, analyzed by flow cytometry. Missing values are due to limited availability in cell numbers required to perform analysis. MN, Mature naive. Mean and SDs are shown. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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