Transfer of RSV-infected <i>Kdm5b</i>-deficient DCs leads to a decreased pathogenic pulmonary RSV challenge.

<p>(A) BMDCs were transfected with <i>Kdm5b</i>-specific siRNA or non-targeting control, then infected with RSV overnight. These BMDCs were transferred intratracheally to the lungs of naïve mice 7 days prior to RSV infection. (B) At 8 days post infection, protein levels were measured from MLN cells restimulated with RSV <i>in vitro</i>. (C) Inflammation around the airways was measured by H&E staining in mice primed with BMDCs, including airway eosinophils (inset). (D) Eosinophils were counted in 100μm sections around the airways. (E) Lungs were dispersed into a single cell suspension using collagenase A and DNase. Esoinophils were measured using flow cytometry. (F) Total lung cells were counted following digestion of lung tissue. CD11c⁺CD11b⁺ and CD11c⁺CD103⁺ DC populations were measured by flow cytometry (G), as well as CD4⁺ T cells and activated CD4⁺CD69⁺ cells (H). (I) RNA was extracted from lung tissue using Trizol reagent, and transcripts of <i>Muc5ac</i>, <i>Gob5</i> and <i>Il13</i> were measured by qPCR. (J) Mucus was visualized in the lungs using PAS staining. Arrows indicate mucus production. (K) Identifying numbers on histology slides were blinded and slides were scored on a scale of 1–4 for mucus production. For all data n = 4-5samples/group and data are representative of two independent experiments. *p<0.05, **p<0.01.