Journal: Nature medicine
Article Title: Adrenergic nerve degeneration in bone marrow drives aging of the hematopoietic stem cell niche
Figure Lengend Snippet: ADRβ3 signaling is essential for maintenance of aging HSCs (a, b) Absolute numbers of MSCs (CD45 − Ter119 − CD31 − CD51 + PDGFRα + ) (a) and HSCs (lineage − Sca-1 + c-Kit + CD48 − CD150 + ) (b) in young and old mice implanted with osmotic pumps containing either saline, the β2-adrenergic agonist clenbuterol or the β3-adrenergic agonist BRL37344 (n=7 young-saline, 6 old-saline, 8 old-clenbuterol and 5 old-BRL37344 mice). (c) Total blood chimerism (CD45.2 + ), blood myeloid chimerism (Mac-1 + CD45.2 + ), blood lymphoid chimerism (B220 + CD4 + CD8 + CD45.2 + ) and HSC chimerism (lineage − Sca-1 + c-Kit + CD48 − CD150 + CD45.2 + ) in CD45.1 recipient mice transplanted with 200 HSCs, derived from the mice described in a and b , at 5 months after transplantation. (n= 5 young-saline, 4 old-saline, 4 old-clenbuterol and 4 old-BRL37344 mice). (d) Bone marrow chimerism following secondary bone marrow transplantation from the mice described in c , 5 months after transplantation. (n=5 young-saline, 4 old-saline, 5 old-clenbuterol and 5 old-BRL37344 mice). (e) . Shown are the Normalized Enrichment Score (NES), p -value and false discovery rate (FDR) of the enrichment. (f) Quantification of mRNA levels of Cxcl12, Scf and Ngf relative to Actb in sorted MSCs derived from mice described in ( a – b ) (n=5 young-saline, 4 old-saline and 4 old-BRL37344; normalized to young-saline). (g) CFU-F frequency from sorted MSCs derived from mice described in a and b (normalized to young-saline; n=5 young-saline, 6 old-saline and 4 old-BRL37344 mice). (h) Absolute numbers of MSCs from Adrb3 +/+ and Adrb3 −/− mice at 2.5 and 5 months of age (normalized to Adrb3 +/+ ; n=4 mice per 2.5-month group, 6 Adrb3 +/+ , 7 Adrb3 −/− per 5 month group). (i, j) CFU-F frequency from sorted MSCs (normalized to Adrb3 +/+ ; n=5 mice per group) (i) and quantification of mRNA levels of Cxcl12, Scf , Angpt1 and Ngf relative to Gapdh in sorted MSCs (normalized to Adrb3 +/+ ; n=5 and 4 Adrb3 +/+ and Adrb3 −/− mice respectively) (j) from 2.5 month old Adrb3 +/+ and Adrb3 −/− mice. (k–n) Absolute numbers of HSCs (k) , LMPPs (lineage − Sca-1 + c-Kit + CD34 + Flt3 + ) (l) , peripheral blood (PB) myeloid cells (Mac-1 + Gr-1 + ) (m) and lymphoid cells (B220 + CD4 + CD8 + ) cells (n) in Adrb3 +/+ and Adrb3 −/− mice at 2.5 and 5 months of age (n=4 mice per 2.5-month group, 7 ( k – i ) and 8 ( m – n ) per 5 month group). Data represented as mean ± s.e.m, p value determined by two-tailed t -test. For box plots, the box spans from the 25th to 75th percentiles and the centerline is plotted at the median. Whiskers represent minimum to maximum range.
Article Snippet: Anti-CD48-PerCp-eFluor710 (HM48-1; 46-0481-182), anti-CD45.1-FITC (A20; 11-0453-85), anti-CD4-PE/Cy7 (GK1.5; 25-0041-82), anti-CD8a-PE/Cy7 (53-6.7; 25-0081-85), anti-B220-APC-eFluor 780 (RA3-6B2; 47-0452-82), anti-CD11b (Mac-1)-APC-eFluor 780 (M1/70; 47-0112-82), anti-Gr-1 (Ly-6G)- APC-eFluor 780 (RB6-8C5; 47-5931-82), anti-CD3e-APC-eFluor 780 (145-2C11; 47-0031-82), anti-CD45-APC-eFluor 780 (30-F11; 47-0451-82), anti-CD31-PE/Cy7 (MEC13.3; 25-0311-82), anti-PDGFRα (CD140a)-APC (APA5; 17-1401-81), anti-PDGFRα (CD140a)-PE/Cy7 (APA5; 25-1401-82), CD51-PE (RMV-7; 13-0512-85), anti-CD41-FITC (MWReg30; 11-0411-81), anti-Ki-67-PerCp-eFluor 710 (SolA15; 46-5698-80), anti-Ter119-APC-eFluor 780 (TER-119; 47-5921-82), Streptavidin APC-eFluor 780 (47-4317-82), anti-CD34-eFluor 660 (RAM34; 50-0341-82 at 1:50 dilution), anti-CD127 (IL7R)-PerCp (A7R34; 45-1271-80), CD135 (Flt3)-PE (A2F10; 12-1351-83) were purchased from eBioscience (Thermo Fisher).
Techniques: Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test