Additional file 1: Figure S1. of Interleukin-21 promotes thymopoiesis recovery following hematopoietic stem cell transplantation

Gating strategies for LSKs. BM cells collected from WT or IL-21R−/− C57BL/6 mice were treated in vitro for proliferation, then stained with the Lin−Sca1+c-kit+ antibody cocktail prior to Ki-67+ incorporation analysis. All described experiments were conducted at least three times with n = 5/group. Figure S2. Functional characterization of T cells. (A) Representative cell trace dilution analysis on CD4+ or CD8+ T cells derived from ctl (unirradiated), PBS-treated, or IL-21-treated LP/J recipient mice. (B) Cytokine quantification by ELISA from T cells derived from the same groups described in panel (A). (C) Representative flow cytometry analysis of granzyme B expression. (D) Quantification of T cells expressing granzyme B. For all presented studies, T cells were stimulated with CD3-CD28 dynabeads for 48 h prior to analyses. All described experiments were conducted at least three times with an n = 5/group. Figure S3. Molecular characterization of T cells. T cells sorted from ctl, PBS-treated, or IL-21-treated LP/J recipient mice were analyzed for their expression of various transcription factors involved in T cell differentiation. All described experiments were conducted at least three times with n = 5/group. Figure S4. Gating strategies for Breg analysis. For detection of IL-10-producing Bregs, CD19+ B cells were first isolated from spleens of treated mice (isotype shown by the filled gray histogram), then stained after in vitro treatment with CD1d and CD5 antibodies. The B cell subset CD1dhiCD5+ was gated prior to IL-10 assessment by intracellular staining. All described experiments were conducted at least three times with n = 5/group. (PDF 2111 kb)