Fig 1: Integrity of EαQ11 nanofibers is required for DC presentation of pEα:I-Ab.(A) Negative-stained TEM images of EαQ11 before (left) and after (right) shearing to reduce nanofiber length. (B) Lengths were quantified by tracing nanofibers using ImageJ software (n = 15 per group). ****P < 0.0001 by unpaired, two-tailed t test. (C) Scheme of experiment. C57BL/6 mice were intranasally (I.N.) immunized with EαQ11. Lung and MLN were collected on day 2 after vaccination. (D) Representative flow cytometry plots displaying pEα:I-Ab–positive CD11b+ or CD103+ DCs in the lung. DCs were identified as CD45+CD49b−TER119−CD19−CD3−SiglecF−Ly6G−CD11c+MHCIIhi cells. pEα:I-Ab–positive DCs were detected using the YAe antibody, which recognizes the pEα:I-Ab complex. The number of pEα:I-Ab–positive APCs in lung (E; black labels) and MLN (F; blue labels). The number of pEα:I-Ab–positive CD11b+/CD103+ DCs in lung (G) and MLN (H). Each dot represents two pooled mice. Data shown are means ± SEM from three independent experiments. ****P < 0.0001 and **P < 0.01 by two-way ANOVA (E to H).