Words similar to podocytes
Example sentences for: podocytes
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RNA and protein samples were analyzed as follows: Podocyte RNA (10 μg) was analyzed by northern blot using probes for HIV-1 nef-LTR [ 7 ] , mouse cyclin D 1 (cDNA generated by RT-PCR from mouse podocytes and verified by sequence analysis), synaptopodin [ 7 ] , and G3PDH.
protocol to study subconfluent podocyte growth and podocyte growth during establishment of mature cell-cell contacts between differentiated podocytes [ 27 28 ] . Briefly, RNA and protein were extracted from parallel plates of podocytes using TRIZOL Reagent or RIPA buffer, respectively, under three conditions: 1) during sub-confluent growth at 33°C; 2) immediately upon reaching confluency (day zero); and 3) on day 14 after switching confluent cultures to 37°C.
[ 11 12 13 14 ] . Kidney transplants between normal and transgenic Tg26 siblings suggested that specific HIV-1 proteins within the kidney, and not circulating factors, transform infected renal epithelium [ 13 ] . Indeed, HIV-1 transgenic mice generated with an HIV-1 proviral construct mutated in the Nef "PXXP" SH3-binding motif for Src-family kinases failed to develop nephropathy [ 31 ] , and a recent in vitro analysis of podocytes infected with single gene mutants of HIV-1 further suggest that Nef plays a central role in causing the proliferative phenotype in HIVAN [ 8 ] . Utilizing these same single gene mutant viruses, we showed here that Nef appears to be play a role in the up-regulation of cyclin D 1 by HIV-1 in vitro . Importantly, these single gene mutants do not exclude the possibility that other HIV-1 gene(s) may cooperate with nef to induce cyclin D1, particularly
These data indicate that HIV-1 expression in infected podocytes induces cyclin D 1 and phospho-pRb (Ser780) expression, hallmarks of cyclin D 1 -mediated G 1 → S progression.
Here, we ask whether infected podocytes expressing HIV-1 genes show the hallmarks of cyclin D 1 -dependent proliferation, specifically the up-regulation of cyclin D 1 transcript and protein expression and the subsequent phosphorylation of pRb on serine 780, a target site of active cyclin D 1 /CDK-4/6 that is not shared with cyclin E/CDK-2 complexes [ 24 ] . Utilizing two well-characterized models of HIVAN pathogenesis, HIV-1 transgenic mice [ 11 12 13 ] and HIV-1 infection of cultured podocytes [ 6 7 8 ] we find not only these hallmarks of cyclin D 1 -dependent proliferation, but also dysregulation of cyclin D 1 expression that normally occurs with cell-cell contact and differentiation [ 25 ] .
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