11.5 2 2 11. RESEARCH ARTICLE RBC ( 1 12 /L) 1.5 1. 9.5 PLT ( 1 9 /L) 1 16 14 HGB (g/l) 19 1 17 16 9. 12 4 4 46 Cellulr & Moleulr Immunology dvne online pulition, PCV (%) 44 MCV (fl) 46 44 ; doi:1.13/mi.214.16 Supplementry Informtion 42 ompnies the pper on Supplementl Figure 4 42 Cellulr & Moleulr4Immunology s wesite (http://www.nture.om/mi). 3 4 Supplementl Figure 5 Supplementl Figure 1 Supplementl Figure Figure 1 Effets 2 of BDE-29 exposure on the hemtologil prmeters in dult mie. BDE-29 exposed mie were intrgstrilly dministered Supplementl BDE-29 Figure with3 different doses of, or mg/kg w t 2-dy intervls. Peripherl lood ws olleted from ontrol nd BDE-29 exposed mie t month 1, nd the hemtologil prmeters inluding red lood ells (RBC), pltelets (PLT), hemogloin (HGB), pked ell volume (PCV) nd men orpusulr volume (MCV) were deteted y HEMAVET 9 multispeies hemtology instrument (DREW Sientifi In.). The dt were nlyzed y one-wy ANOVA (men6s.d.). BDE, rominted diphenyl ether; w, ody weight. Cellulr & Moleulr Immunology: doi:1.13/mi.214.16
11.5%.5% 77.9% 79.% 1.% 13.5% 1.3% 1.1% CD4 2.9% 3.2% 2.% 2.4% CD % CD4 SP 2 15 1 5 % CD SP 5 4 3 2 1 % CD4CD DP 9 5 75 7 65 # of CD4 SP/thymus ( 1 6 ) 3 2 1 # of CD SP/thymus ( 1 6 ) 6 4 2 # of CD4CD DP/thymus ( 1 6 ) 1 # thymoytes/thymus ( 1 6 ) 2 1 Supplementl Figure 2 BDE-29 exposure did not signifintly ffet the perentges nd numers of T-ell supopultions in the thymus. Thymoytes were olleted from ontrol nd BDE-29 exposed mie with dose of, or mg/kg w t month 3, nd the perentges nd numers of CD4 1 single-positive (CD4 SP), CD 1 single-positive (CD SP) nd CD4 1 CD 1 doule-positive (CD4CD DP) T ells in thymus were lulted y flow ytometry through stining with nti-cd3e, nti-cd4 nd nti-cd. () As shown in the flow digrm (gted on thymoytes), the perentges indite the popultion of CD4 SP, CD SP nd CD4CD DP T ells in thymoytes. The dt were nlyzed y one-wy ANOVA (men6s.d.). BDE, rominted diphenyl ether; w, ody weight. Cellulr & Moleulr Immunology: doi:1.13/mi.214.16
.2%.2%.1%.1% CD11 Ly6G CD11.1%.1%.1%.1% % Neu in thymoytes # of Neu/thymus ( 1 6 ).4.3.2.1..4.3.2.1. CD11 % DCs in thymoytes # of DCs/thymus ( 1 6 ).2.15.1.5..2.15.1.5. Supplementl Figure 3 BDE-29 exposure did not ffet the perentges nd numers of CD11 1 Ly6G 1 neutrophils nd CD11 1 CD11 1 dendriti ells in the thymus. Thymoytes were isolted from ontrol nd BDE-29 exposed mie with dose of, or mg/kg w t month 3, nd the perentges nd numers of CD11 1 Ly6G 1 Neu nd CD11 1 CD11 1 DCs were lulted y flow ytometry through stining with nti- CD11, nti-ly6g nd nti-cd11. () As shown in the flow digrm (gted on thymoytes), the perentges (%) indite the popultion of CD11 1 Ly6G 1 neutrophils nd CD11 1 CD11 1 dendriti ells in thymoytes. The dt were nlyzed y one-wy ANOVA (men6s.d.). BDE, rominted diphenyl ether; w, ody weight; DC, dendriti ell; Neu, neutrophils. Cellulr & Moleulr Immunology: doi:1.13/mi.214.16
IL-2 IFN-g BDE-29 27. 7. CD44 27.3 9.2 % IFN-g of CD + T % IL-2 of CD + T 4 3 2 1 3 2 1 ** * SSC BDE-29 Supplementl Figure 4 Long-term exposure to BDE-29 y dose of mg/kg w hd dverse effets on the funtionlity of spleni CD T ells. Splenoytes were olleted from ontrol nd BDE-29 exposed mie t month 27, nd IFN- nd IL-2 produtions of CD T ells were mesured y intrellulr ytokine stining following PMA ( ng/ml) nd ionomyin ( ng/ml) stimultion for 5 h in vitro. As shown in the flow digrm (gted on CD 1 T ells), the perentges (%) indite IFN-- or IL-2-positive popultion of CD 1 T ells (left pnels). Br grphs disply the orresponding sttistil results of IFN- 1 CD 1 T ells or IL-2 1 CD 1 T ells etween ontrol nd BDE-29 exposed mie. The dt were nlyzed y using unpired Student s t-test (men6s.d., *P,.5, **P,.1). BDE, rominted diphenyl ether; w, ody weight. Cellulr & Moleulr Immunology: doi:1.13/mi.214.16
T CM 16.3 BDE-29 14.4 T EM CD27 CD27 CCR7 3.7 2.6 141 123 CCR7 12.4.9 BDE-29 12 19 6. CD44 5. CD122 CD127 Supplementl Figure 5 Effet of BDE-29 exposure on the phenotype of CD T ells. BDE-29 exposed mie were intrgstrilly dministered with BDE-29 y dose of mg/kg w t 2-dy intervls, nd the peripherl lood were olleted from ontrol nd BDE-29 exposed mie t month 5. () The expressions of CD27 nd CCR7 were nlyzed on the entrl memory (T CM,CD44 hi CD62L hi ) or effetor memory (T EM, CD44 hi CD62L lo ) phenotype CD T ells. () The perentges (%) indite the popultions of CD44 hi CD27 hi, CD44 hi CD27 lo, CD44 hi CCR7 hi nd CD4 hi CCR7 lo of CD 1 T ells. () The numers indite the MFI of CD122 nd CD127 on CD 1 T ells. BDE, rominted diphenyl ether; w, ody weight; MFI, medin fluoresent intensity. Cellulr & Moleulr Immunology: doi:1.13/mi.214.16