Mastitis induces SEMA5A through FEZL. (A) FEZL expression. Results are means of quantity relative to brain. Green bars indicate the mean (n = 3) from the same cattle, and red and blue bars indicate the mean (n = 3) from other cattle with mastitis. The orange bar indicates the mean ± SEM (n = 54) from 18 randomly collected cattle with unknown status. Symbols denote statistically significant results as determined by Student’s t test (∗∗, P < 0.0005; ∗∗∗, P < 0.0001). (B) Luciferase assay of FEZL with SEMA5A1 (red) and SEMA5A2 (blue). Results are means ± SEM of relative activity to COS7 cells transfected without FEZL. Asterisks denote statistically significant results as determined by Student’s t test (∗∗, P < 0.01; ∗∗∗, P < 0.005). (C) SEMA5A expression in OCUB-M cells transfected with FEZL. Results are means ± SEM (n = 9; three experiments) of quantity relative to OCUB-M cells transfected without FEZL after adjusting for the FEZL expression level. Asterisks denote statistically significant results as determined by Student’s t test (∗, P < 0.05; ∗∗∗, P < 0.005).