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. Author manuscript; available in PMC: 2019 Jan 23.

Published in final edited form as: Mucosal Immunol. 2018 Jul 23;11(5):1524–1536. doi: 10.1038/s41385-018-0052-1

Fig. 4 — DLL4 inhibition changed Th cytokine profile in vivo during RSV infection. a Wild-type Balb/c mice were intratracheally infected with Line 19 RSV. DLL4 expression were measured at 6 days post infection (dpi). N = 4 in each group. b DLL4 were neutralized at 0, 2, 4, 6 dpi. Periodic acid-Schiff (PAS) staining of formalin-fixed lung section from 8 dpi. Bar, 200 μm. ↑ indicates the detection of mucin, and designates mononuclear cells aggregates. c Smyd3 expression in mediastinal lymph node (mLN) were measured at 6 dpi after DLL4 inhibition. N = 10 in each group. d Foxp3-EGFP knock-in mice were intratracheally infected with RSV. DLL4 were neutralized at 0, 2, 4 dpi. mLN cells were collected. Viable CD4 + Foxp3-EGFP+/− cells were sorted from mLN and Smyd3 expression were measured. N = 3 in each group. e Viable CD4 + Foxp3-EGFP+/− cells were sorted from mLN. Foxp3 mRNA were measured. N = 3 in each group. f Il5 mRNA in both Foxp3-EGFP− and Foxp3-EGFP+ were measured. g Ifng mRNA in both Foxp3-EGFP− and Foxp3-EGFP+ were measured. h Foxp3-EGFP knock-in mice were intratracheally infected with RSV. DLL4 were neutralized at 0, 2, 4, 6 dpi. mLN cells were collected at 8 dpi. Viable CD4 + Foxp3-EGFP+/− cells were sorted from mLN, and Il17a expression in both Foxp3-EGFP− and Foxp3-EGFP+ were measured. i Il10 expression in both Foxp3-EGFP− and Foxp3-EGFP+ viable CD4 T cells in mLN were measured at 8 dpi. j IL-10 production by control IgG or anti-DLL4-treated mLN cells were harvest at 8 dpi and measured after 48 h of RSV re-stimulation. Data represent mean ± SEM. Data were from one experiment representative of two experiments with 3~10 mice per time point, with samples from each mouse processed and analyzed separately. * P < 0.05; ** P < 0.005; *** P < 0.0005; NS, no significance (unpaired two-tailed t-test)

Inline graphic — DLL4 inhibition changed Th cytokine profile in vivo during RSV infection. a Wild-type Balb/c mice were intratracheally infected with Line 19 RSV. DLL4 expression were measured at 6 days post infection (dpi). N = 4 in each group. b DLL4 were neutralized at 0, 2, 4, 6 dpi. Periodic acid-Schiff (PAS) staining of formalin-fixed lung section from 8 dpi. Bar, 200 μm. ↑ indicates the detection of mucin, and designates mononuclear cells aggregates. c Smyd3 expression in mediastinal lymph node (mLN) were measured at 6 dpi after DLL4 inhibition. N = 10 in each group. d Foxp3-EGFP knock-in mice were intratracheally infected with RSV. DLL4 were neutralized at 0, 2, 4 dpi. mLN cells were collected. Viable CD4 + Foxp3-EGFP+/− cells were sorted from mLN and Smyd3 expression were measured. N = 3 in each group. e Viable CD4 + Foxp3-EGFP+/− cells were sorted from mLN. Foxp3 mRNA were measured. N = 3 in each group. f Il5 mRNA in both Foxp3-EGFP− and Foxp3-EGFP+ were measured. g Ifng mRNA in both Foxp3-EGFP− and Foxp3-EGFP+ were measured. h Foxp3-EGFP knock-in mice were intratracheally infected with RSV. DLL4 were neutralized at 0, 2, 4, 6 dpi. mLN cells were collected at 8 dpi. Viable CD4 + Foxp3-EGFP+/− cells were sorted from mLN, and Il17a expression in both Foxp3-EGFP− and Foxp3-EGFP+ were measured. i Il10 expression in both Foxp3-EGFP− and Foxp3-EGFP+ viable CD4 T cells in mLN were measured at 8 dpi. j IL-10 production by control IgG or anti-DLL4-treated mLN cells were harvest at 8 dpi and measured after 48 h of RSV re-stimulation. Data represent mean ± SEM. Data were from one experiment representative of two experiments with 3~10 mice per time point, with samples from each mouse processed and analyzed separately. * P < 0.05; ** P < 0.005; *** P < 0.0005; NS, no significance (unpaired two-tailed t-test)