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. 2017 Oct 23;8:1346. doi: 10.3389/fimmu.2017.01346

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Copyright © 2017 Ichii, Kimura, Okamura, Horino, Nakamura, Sasaki, Elewa and Kon.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Protein expression and localization of IL-36α in unilateral ureter obstruction (UUO) kidneys. (A) Renal histopathology after UUO. In periodic acid Schiff staining, dilated distal tubules (DTs) are observed from 0.5 h after UUO (arrows). Positive immunostaining for calbindin D28k, a DT marker, is observed in DTs, and almost all of them were dilated from 0.5 h after UUO (arrows). IL-36α⁺ immunostaining is observed in DTs, especially in the macula densa (MD), from 12 h after UUO (arrows). This immunostaining spreads to the entire DTs at 24 h after UUO. (B) Immunohistochemistry for IL-36α in the UUO kidney at 24 h after UUO. IL-36α⁺ immunostaining is observed in the cytoplasm, and some nuclei are also stained. (C) Immunohistochemistry for IL-36α in the UUO kidney at day 11 after UUO. IL-36α⁺ tubules are numerous in UUO kidneys compared to Cont kidneys. (D) The number of IL-36α⁺ tubules in Cont and UUO kidneys. Values = mean ± SE. A significant difference from the Cont in same sexes is indicated by *(P < 0.05) or **(P < 0.01). No sex-related difference is detected. n = 5 (kidneys). (E) Time course of IL-36α⁺ tubule number in Cont and UUO kidneys. (F) The number of IL-36α⁺ tubules in Cont and UUO kidneys. Values = mean ± SE. A significant difference from the Cont is indicated by *(P < 0.05) or **(P < 0.01). A significant difference from 12 h in the same group is indicated by ^†(P < 0.05) or ^††(P < 0.01). n ≥ 5 kidneys. (G) Immunohistochemistry for IL-36α⁺ tubules in Cont kidney. In a few Cont kidneys, IL-36α⁺ immunostaining is occasionally observed in the MD at 24 h after UUO (arrows). (H) Probability of IL-36α⁺ tubules in DT segments attached to the renal corpuscles and the other DT segments. Values = mean ± SE. A significant difference from others is indicated by *(P < 0.05) or **(P < 0.01). A significant difference between Cont kidneys and UUO kidneys in the same group is indicated by ^†(P < 0.05) or ^††(P < 0.01). n = 5 (kidneys). (I) Immunofluorescence for calbindin D28k and IL-36α. No IL-36α⁺ staining is detected in Cont kidney. In UUO at 24 h, dilated calbindin D28k⁺ tubules (green, large arrow) are also positive for IL-36α (yellow, arrowheads), and some of them show decreased calbindin D28k⁺ staining (red, small arrows).