Supplemental Figure S8.

Protein kinase A (PKA) inhibitor H-89 blocks 8-Br-cAMP–mediated cystogenesis and Creb phosphorylation in proximal tubules. A and B: Representative stereomicroscopic images of kidney explants cultured for 5 days (120 hours) in media containing 100 μmol/L 8-Br-cAMP or 100 μmol/L 8-Br-cAMP + 10 μmol/L H-89. Cysts are present in explants treated with 8-Br-cAMP alone (concave arrowhead), but are absent in the 100 μmol/L 8-Br-cAMP + 10 μmol/L H-89 treatment group (B). C and D: Representative hematoxylin and eosin images of sections obtained from kidney explants cultured with 100 μmol/L 8-Br-cAMP or 100 μmol/L 8-Br-cAMP + 10 μmol/L H-89 at 120 hours. Cysts (concave arrowhead) can be seen in explants treated with 8-Br-cAMP alone (C), but are absent in the 100 μmol/L 8-Br-cAMP + 10 μmol/L H-89 treatment group (D). E and F: Representative co-immunofluorescence images for p-Creb (red) and lotus tetragonolobus lectin (LTL) (green) on sections obtained from kidney explants cultured with 100 μmol/L 8-Br-cAMP or 100 μmol/L 8-Br-cAMP + 10 μmol/L H-89 for 5 days. In 8-Br-cAMP–treated explants, p-Creb is present in cysts with reduced LTL staining (E, concave arrowheads); however, in the 100 μmol/L 8-Br-cAMP + 10 μmol/L H-89 treatment group, p-Creb signal (F, concave arrowheads) largely is absent in LTL-positive tubules. Arrowheads indicate p-Creb staining in nephron progenitors. Scale bars: 500 μm (A and B); 100 μm (C and D); 50 μm (E and F).