Relationships between renal cytoplasmic and nuclear aldosterone-receptors

Relationships between renal cytoplasmic and nuclear aldosteronereceptors.Three 3H-aldosterone receptor complexes have been recovered from rat kidneys: 1) cytosol (high speed supernatants), 2) Tris-soluble nuclear (obtained by an osmotic shock procedure), and 3) chromatin-bound (prepared by extracting post-shock nuclei with 0.4 M KCl).Glycerol density gradient analyses of cytosol labelled in vivo or in vitro with 3H-aldosterone yielded two specific peaks -4.5S and 8.5S.These peaks were sensitive to salt concentration; 0.4 M KCl shifted the 8.5S to 4.5S and the addition of Ca++ (6 mM) resulted in a further shift to 3.5S.The Tris-soluble nuclear species sedimented at 3S and the chromatin-bound species at 4S.The time-course of generation of the 3H-aldosterone-labelled cytosol and nuclear receptor species was studied in vivo and in vitro by tissue slice and reconstitution methods.The results obtained are consistent with a three-step mechanism: cytosol (8.5S or 4.5S)→ Tris-soluble nuclear (3S)→ chromatin-bound (4S).Alternatively, the 3S and 4S complexes may be attached to independent nuclear sites.The formation of the chromatin-bound species was temperature sensitive and failed to form at 0°C.Pre-treatment with DNase but not RNase impaired the generation of both the Tris-soluble nuclear and chromatin-bound species.These results imply a close association between nuclear aldosterone-receptor complexes and intact DNA

Anti-oxidant NAC attenuated the decrease in fitness caused by FLT.

<p>Fitness was measured by the number of sigmoidal body bends per worm per minute. FLT concentration = 200μM. Anti-oxidant concentration = 100μM. Statistics were calculated with a two way ANOVA with Turkey’s multiple comparisons test from 10 worms per treatment condition over ≥3 independent trials. FLT compared to control of that same time point. FLT + NAC compared to FLT of that same time point. * = P<0.05, ** = P<0.01, *** = P<0.001, n.s. = not significant.</p

mtDNA copy numbers fluctuated and eventually became normalized during continued short-term exposure (6h).

<p>Relative quantities (%) of mtDNA compared to control animals. Numbers between parentheses indicate 95% confidence intervals (51df) from ≥3 replicates per 2 individual experiments. Significance was determined using a two-tailed student’s <i>t</i>-test assuming unequal variance compared to control animals. * = P-value <0.05, ** = P-value <0.01, *** = P-value <0.001.</p

NRTIs caused average lifespan extension.

<p>Lifespan extension is dependent on the timing of exposure: A, exposure from larval L1; B, exposure from larval L4. NRTI concentration = 200μM. All animals are N2 at 20°C, mean and maximum refer to the amount of days after L1 (A), and L4 (B). Statistics of NRTIs at L4 were conducted compared their respective DMSO control (AZT, d4T, ddI, & FLT = 0.067%; ddC = 0.2%). All other statistics are compared to controls. SEM = standard error of the mean.</p

Overlap of thymidine analogue induced DEGs with genes known to be regulated by HIF-1, CEP-1 and SKN-1 after 72h exposure.

<p>The expression direction (Up or Down) of thymidine analogue DEGs, with an adjusted P-value <0.01, at 72h, are compared to respectively Up or Down regulated genes from the different conditions taken from [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0187424#pone.0187424.ref044" target="_blank">44</a>–<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0187424#pone.0187424.ref046" target="_blank">46</a>]. The numbers of genes that overlap per condition at each time point are given with their representative percentage of thymidine analogue regulated genes between brackets. Over-represented P-value: * = P<0.05, ** = P<0.01, *** = P<0.001, n.s. = not significant. n.a. = not applicable.</p

NAC can mitigate the average lifespan increase caused by FLT.

<p>All animals are N2 exposed from L4 at 20°C. Statistics were conducted compared to the DMSO control. FLT concentration = 200μM, NAC concentration = 100μM.</p

NRTIs reduced the number of thrashes per worm per minute.

<p>NRTI concentration = 200μM. Statistics were calculated by a two way ANOVA with Dunnett’s multiple comparisons test, compared to the control of that same time point, of 10 worms per treatment condition over ≥3 independent trials. * = P<0.05, ** = P<0.01, *** = P<0.001, n.s. = not significant.</p

NRTIs reduced nematode body length.

<p>Relative body length was measured after 48hrs NRTI exposure of L1 and 96h exposure of L4 animals (30 worms in ≥2 individual experiments). NRTI concentration = 200μM. Statistics were calculated with a two-way ANOVA with replication, compared to controls. *** = P<0.001.</p