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. 2018 Jul 16;9:1589. doi: 10.3389/fimmu.2018.01589

Figure 4.

Figure 4

Expression of P2YR subtypes on whole kidney tissue, isolated glomerula, isolated podocytes versus non-podocyte glomerular cells, and blood leukocytes following nephrotoxic serum (NTS)-induced glomerulonephritis (GN). C57BL/6 mice received either vehicle (PBS) or NTS (solved in PBS) intravenously on days 1, 2, and 3 to induce GN. For the isolation of podocytes, we used Gt(ROSA)26Sortm4(ACTB-tdTomato,-EGFP)Luo/J mice, which had been crossed with hNPHS2Cre mice. Podocytes of these mice express GFP while all other cells express Tomato. On days 4, 6, and 10 animals were killed after a blood sample for the isolation of leukocytes had been collected. RNA was isolated from whole kidney tissue, isolated glomerula, isolated podocytes, and non-podocytal glomerular cells and from blood leukocytes. Glomerula were isolated using perfusion with magnetic beads. From the isolated glomerula, we separated the GFP-labeled podocytes from non-podoyctes via FACS. Relative expression of the different P2Y receptors compared to β2m in triplicates were analyzed using quantitative RT-PCR. (A) Expression of P2YR subtypes on whole kidney tissue from n = 5 animals per group. (B,C) Expression of P2YR subtypes on isolated glomerula from n = 5 animals per group. (D) Expression of P2YR subtypes on isolated podocytes versus non-podocyte glomerular cells from n = 5 animals per group. (E) Expression of P2YR subtypes on isolated blood leukocytes from n = 5 animals per group. Data are shown as mean ± SEM, n = 5 mice in each group. Data from one representative experiment out of three are shown.