Fig. 6

Macrophage autophagy-related 5 (Atg5) deletion impaired the CCR6-CCL20 axis after I/R and UUO injury. (A and B) Flow cytometric analysis determined that F4/80⁺ macrophage infiltration injured kidneys at 1, 2, 3, 4, 5, and 9 days after undergoing 31-min unilateral renal ischemia/reperfusion (UI31R). (C, D and E) MΦ atg5^−/− and WT mice were sacrificed 4 days after UI31R injury. (C) F4/80⁺ macrophages isolated from kidney single-cell suspensions from MΦ atg5^−/− and WT mice using immunomagnetic positive selection. Flow cytometric analysis identified the isolated cell purity. (D and E) The renal macrophage mRNA levels of Atg5, TNFα, Il6, CCR6 and CXCR2 in MΦ atg5^−/− and WT mice. Data are presented as the mean ± SEM. ** P < 0.005, **** P < 0.0001 (n = 3–4 per group). Unpaired, two-tailed Student’s t test. (F) The renal mRNA level of CCL20 was significantly lower in MΦ atg5^−/− mice than in WT mice at 4 and 28 days after UI31R injury and 14 days after UUO injury. Data are presented as the mean ± SEM. **** P < 0.0001 (n = 3 per group). Two-way ANOVA with Sidak’s multiple-comparison test