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. 2016 Jul 20;311(3):F626–F639. doi: 10.1152/ajprenal.00196.2016

Fig. 8.

Fig. 8.

Immunostaining for PEC activation marker CD44 and collagen IV is higher in PECs in APD mice given rapamycin. A–D: confocal microscopy showing immunofluorescent double staining for CD44 (red, solid arrow) and PAX8 (green, arrowheads) (magnification: ×400). PAX8 staining was used to demarcate PECs. Scale bars are provided. Nuclei were stained blue with 4′,6-diamidino-2-phenylindole (DAPI). A: representative image of glomeruli at baseline in which CD44 expression in glomeruli is absent. A′: a PEC negative for CD44 is seen magnified and is shown as single-channel images below. B: expression of CD44 increases in PECs at D14 APD, as indicated by solid arrow. PEC surrounded by CD44 is shown magnified in B′ and is shown as single channels below. C: CD44 expression is higher in PECs following rapamycin treatment. The dashed box demonstrating CD44 in PEC on Bowman's capsule is shown in C′ and as single images to the right. D: graph showing percentage of glomeruli positive CD44 at baseline, D14, and D14 + rapamycin, with increasing number of glomeruli positive at D14 APD and again significantly higher at D14 APD + rapamycin. Scale bars are provided. Nuclei were stained blue with DAPI. E–H: confocal microscopy (magnification: ×400) showing double immunofluorescent staining for collagen IV (red, dashed arrow) and PAX8 (arrowhead) expression in glomeruli at baseline (E). Dashed arrow indicates positive staining along Bowman's capsule at baseline, which is delicate and thin, consistent with quiescent PECs. Collagen expression is adjacent to PAX8+ PEC (dashed box) and is shown in E′ and as single-channel images to the right. F: expression of collagen IV increases in PECs at D14 APD, as indicated by arrowhead and dashed arrow. Area in dashed box is shown in F′ and as single-channel images to the right. G: collagen IV expression is higher in PECs following rapamycin treatment. The area positive for collagen IV and PEC in the dashed box is shown in G′ and as single-channel images to the right. H: graph showing area positive along Bowman's capsule for collagen IV increases from baseline compared with D14 and D14 + rapamycin. Scale bars are provided. Nuclei were stained blue with DAPI.