Supplemental Figure 1

Supp. Figure 1. Effects of aging on pulmonary architecture and inflammatory cell dynamics in O3 induced lung injury. (A) Representative hematoxylin and eosin stained tissue showing low magnification visualization of a 12 wk (young) and 80 wk (old) SP-C^I73T lung 24 h after O₃ exposure (3 h, 0.8ppm). (B) Blood counts from juvenile/young adults (8-14 wk), middle aged (44-52 wk), and old (>80 wk) mice after exposure to filtered air (control) or 24 h and 72 h O₃ (3 h, 0.8ppm). Analysis was performed on SP-C^WT and SP-C^I73T mice. Data are presented as mean ± SE (n = 3-14 mice/group), analyzed using two-way ANOVA. A p<0.05 was considered significant. (C) Stacked column graph combining relative abundance of SigF+CD11c+CD11b- alveolar macrophages, CD11c⁺CD43⁺ interstitial macrophages, CD11b⁺CD11c^- monocytes, and CD11b⁺CD43^-CD11c⁺ monocyte-derived macrophages (MoDMs) from juvenile/young adults (8-14 wk) and old (>80 wk) SP-C^WT and SP-C^I73T mice. (D) Relative abundance of CD8⁺CD103⁺ dendritic cells from juvenile/young adults (8-14 wk) and old (>80 wk) SP-C^WT and SP-C^I73T mice. (E) Stacked column graph combining relative abundance of B220+CX3CR1- and B220⁺CX₃CR1⁺ B cells from juvenile/young adults (8-14 wk) and old (>80 wk) SP-C^WT and SP-C^I73T mice. Data are presented as mean ± SE (n = 3-9 mice/group), analyzed using three-way ANOVA. ^a Significant difference from SP-C^WT mice; ^b Significant difference from young mice. ^c Significant difference from air control mice. d Significant difference from 24 h O₃. A p<0.05 was considered significant. (F) Luminex analysis of immunoglobulin M (IgM) in BAL fluid from juvenile/young adults (8-14 wk), middle aged (44-52 wk), and old (>80 wk) mice after exposure to filtered air (control) or O₃ (3 h, 0.8ppm). Ozone time points (24 h and 72 h) have been combined. Data are presented as mean ± SE (n = 4-17 mice/group), analyzed using three-way ANOVA. A p<0.05 (*) was considered significant. (G) Representative immunofluorescent image of B220⁺ staining of B cells within tertiary lymphoid tissue in O₃ exposed old SP-C^I73T mouse. B220 is shown in green (AlexaFluor488), DAPI was used to identify cell nuclei. (H) Representative immunohistochemical staining of CD19⁺ B cells within tertiary lymphoid tissue in O₃ exposed old SP-C^I73T mouse.