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. 2022 Oct 17;119(43):e2123187119. doi: 10.1073/pnas.2123187119

Fig. 2.

Fig. 2.

Disruption of AEC2 proteostasis by SftpcC185G promotes early lung injury in mice. (A) Weight change following IP TMX to SftpcWT and SftpcC185G mice (n = 10 and 7, respectively). (B) Kaplan–Meier survival curve SftpcWT and SftpcC185G mice (n = 10 and 6, respectively), P = 0.0012 by log-rank (Mantel–Cox) test. (C) Representative hematoxylin and eosin (H&E) staining of lung section 7 d after TMX shows alveolitis in SftpcC185G lungs (top 4× magnification, scale bars = 1,000 μM; bottom 10× magnification, scale bars = 100 μM). (D and E) BALF cell counts (D) and percentage of BALF cells (E) that are granulocytes by Giemsa staining in SftpcWT and SftpcC185G mice at 7 d after TMX. ***p < 0.0005, ****p < 0.0001 by two-way t test. (F) Schematic for IP bortezomib (BTZ) in vivo proteasome inhibition studies (vehicle = 0.1% dimethyl sulfoxide [DMSO] in saline) in SftpcWT and SftpcC185G mice receiving OG TMX. (G) Western blotting for polyubiquitin (Ubq), BIP, and ProSP-C in AEC2 lysates isolated 6 h after in vivo BTZ (or vehicle) treatment. (H) (Top) Representative Immunostaining of lung section from SftpcC185G lungs at 4 and 7 d after BTZ or vehicle for proSP-C (20× magnification scale bars = 50 μM) and (Bottom) AEC2 mean fluorescence intensity (MFI) for proSP-C as measured in 40 most fluorescent AEC2s per 20× field per mouse (three 20× fields assessed per mouse). Not significant (ns), *p < 0.05, **p < 0.005, ***p < 0.0005, ****p < 0.0001 by one-way ANOVA with Tukey’s multiple comparisons test. (I) Mean weight change from baseline following OG TMX to SftpcC185G mice followed by BTZ or vehicle treatment. Data are shown as mean ± SEM (vehicle n = 5, BTZ n = 7). (J) Kaplan–Meier survival curve of SftpcC185G mice with OG TMX followed by BTZ or vehicle treatment (vehicle n = 5, BTZ n = 7) P = 0.0082 by log-rank (Mantel–Cox) test. (K and L) BALF cell counts (K) and percentage of BALF cells (L) that are granulocytes by manually counting Giemsa-stained cytospins of SftpcC185G mice at 7 d after OG TMX followed by BTZ or vehicle treatment. **p < 0.005, ***p < 0.0005 by two-way t test.