Words similar to podocyte
Example sentences for: podocyte
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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.
Following 7.5% SDS/PAGE, podocyte protein (20 μg) was analyzed by western blot for cyclin D 1 (ECL detection of a 1:1000 dilution of rabbit anti-cyclin D 1 antibody, Santa Cruz BioTechnology).
Cyclin D 1 is characterized as the major D-type cyclin controlling G 1 -phase progression in kidney cells, both during normal and abnormal proliferation [ 22 23 ] . To determine whether cyclin D 1 may be active in promoting G 1 → S progression in kidneys expressing HIV-1 genes, we utilized a well-characterized HIV-1 transgenic mouse model (Tg26) of HIVAN [ 11 12 13 ] . Secondary to renal expression of the HIV-1 NL4-3 Δ gag-pol proviral transgene, adolescent Tg26 transgenics develop a progressive, proliferative renal disease marked by podocyte and tubular epithelial hyperplasia that is indistinguishable both clinically and histopathologically from human HIVAN.
After the 14-day podocyte differentiation protocol described above, total RNA was extracted from wild-type and HIV-1-infected podocytes before treatment with flavopiridol (Developmental Therapeutics Program, National Cancer Institute) (day 14), two days after suppression of HIV-1 transcription with media containing 50 nM flavopiridol (day 16), and one day after drug washout using media without flavopiridol (day 17).
In contrast, cell-cell contact in wild-type and control-infected, but not HIV-1-infected podocytes, caused a severe down-regulation of cyclin D 1 transcript and protein concomitant with the up-regulation of synaptopodin, a podocyte differentiation marker [ 7 28 ] , by the end of the 14-day differentiation period.
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