Fig. 4
P2X7 is upregulated in renal tubule cells exposed to TGF-β1 and in patients with CKD and in the UUO mouse mdoel. Staining of biopsy material isolated from individuals with DN was performed for P2Y2 (a & b), P2Y6 (f & g), and P2X7 (k & l). Staining for non-diabetic controls can be observed in P2Y2 (c & d), P2Y6 (h & i), and P2X7 (m & n). Magnification: 400X. hPTECs were also cultured in low glucose with/without TGF-β1 (2-10 ng/mL) for 48 h. Whole-cell abundance of P2Y2 (p), P2Y6 (q) and P2X7 (r) were determined through densitometry. Representative blots for each protein are shown, with expression normalized by re-probing for ɑ-tubulin as a loading control. Bars correspond to their associated lanes in the respective blot. qRT-PCR was performed on tubules isolated from WT (UUO) and Cx43+/− (UUO) for P2Y2 (s), P2Y6 (t) and P2X7 (U). qRT-PCR are expressed in graphs as arbitrary units (a.u.) that represent the ratio of the target gene to internal control gene (HPRT). Results were from three or more separate experiments; with key significance shown: *P < 0.05; ***P < 0.001