Fig. 3
From: HIV-1 gp120-Induced Tubular Epithelial Cell Apoptosis Is Mediated Through p38-MAPK Phosphorylation
(A) Western blotting for renal cortical tissue CD4 detection. Human renal cortical tissue was homogenized in RIPA buffer with protease inhibitors. Subsequently, 10, 25, or 50 µg of protein/lane was loaded, separated on PAGE, immunoprecipitated with specific antibody for CD4, immunoblotted with HRP-labeled anti-rabbit IgG, and detected by ECL. All samples showed a band of 58-kDa protein. (B) Western blotting for tubular cell CD4 detection. HGEC, human T cells (Jurkat cells, JKT), mouse macrophage cell line (J774), and human monocyte cell line (U937) were used as positive controls; normal rat kidney (NRK) cells were used as a negative control. Human proximal renal tubular cells (HK2) and other cells were grown to subconfluence, cells were lysed, protein extracted, Western blots generated and probed for CD4. HK2, HGEC, JKT, J774, and U937 cells show a 58-kDa band, whereas NRK did not show 58-kDa band. (C) Tubular cell mRNA expression of CD4. A representative Northern blot showing mRNA expression of CD4 receptors by human proximal tubular epithelial cells (lane 2). Similarly, glomerular epithelial cells (lane 1), U937 (lane 3), and Jurkat cells (lane 4) showed mRNA expression of CD4. However, human lung fibroblasts (negative control, lane 5) did not show CD4 mRNA expression.