Glomerular morphology was disrupted in adriamycin treated mice.

<p>Kidneys were collected for morphological analysis at day 12 after adriamycin induced nephropathy. Slides were blindly examined by a pathologist (n = 15 for all groups). Shown are representative images of (A) controls, which present a normal glomerular structure and foot processes, (B) untreated adriamycin mice, displaying a high degree of foot process effacement, (C) treatment with BMS-470539 or (D) α-MSH, showing disrupted glomerular structures similar to untreated adriamycin mice. (E) The number of podocyte foot processes per 10 µm of glomerular basement membrane was quantified. Untreated adriamycin (AD) and mice treated with BMS-470539 (BMS) or α-MSH had a lower number of foot processes compared to controls (Ctrl). *Normal foot process, → disrupted glomerular barrier structure and loss of foot processes. Scale bar = 2 µm. FP = foot processes, GBM = glomerular basement membrane.</p

MC1R protein expression.

<p>Western blotting was performed in order to detect MC1R protein expression in mouse and human tissue. Left panel: incubation with a rabbit polyclonal MC1R antibody (Alomone Labs). Right panel: incubation with MC1R antibody and control blocking peptide antigen (BP) in order to show antibody specificity. A) mouse whole glomerular lysate, B) cultured wild type mouse podocytes and C) a human malignant melanoma cell line A375. MC1R protein expression was detected at the expected size of 37 kDa (left panel).</p

Overview of the results.

<p>*Values in parenthesis indicates time after ^99mTc-DTPA injection where highest deviation from control was observed</p><p>ND = not done</p><p>Overview of the results concerning body weight, blood cell count, urea and creatinine blood levels, ^99mTc-DTPA and ^99mTc-DMSA scintigraphy, and histology. The results are presented as test divided by control (given as percent)</p

Urinary bladder content following ^99mTc-DTPA administration.

<p>Urinary bladder content at 27.5 min after ^99mTc-DTPA administration. Scintigraphy was performed at 4, 8, and 12 months after administration of 30–150 MBq ¹⁷⁷Lu-octreotate. Kidney uptake is presented as percent of injected activity. Error bars represent SEM</p

Upstream regulators.

<p>Top upstream regulators of transcripts with recurrent expression (cf. Figs <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0136204#pone.0136204.g002" target="_blank">2</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0136204#pone.0136204.g003" target="_blank">3</a>) in kidney cortex and medulla</p

MC1R agonists did not reduce albuminuria in adriamycin-treated mice.

<p>(A) Five different doses of adriamycin, in the range of 5–10 mg/kg, were intravenously injected and the level of albuminuria was measured after 7 days. The urinary albumin-to-creatinine ratio (UACR) increased with increasing levels of adriamycin. (B) Albuminuria at day 7. Treatment with the MC1R agonist BMS-470539 did not reduce the level of albuminuria for the 8 mg/kg adriamycin dose, on the contrary it slightly increased albuminuria for the 10 mg/kg dose (n = 17–19, p<0.05). The unspecific melanocortin receptor agonist α-MSH did not have any significant effect (n = 9, n.s.) compared with untreated adriamycin mice. Results are presented as mean ± SEM.</p

Number of differentially regulated transcripts.

<p>Number of differentially regulated transcripts in (a) kidney cortex and (b) kidney medulla 4, 8 and 12 months after administration of 30–150 MBq ¹⁷⁷Lu-octreotate. Distribution of up- and down-regulated transcripts is indicated with positive and negative numbers, respectively</p

Recurrently regulated transcripts in kidney medulla.

<p>Gene expression patterns for differentially regulated transcripts with recurrent expression at one or more time points in kidney medulla 4, 8, or 12 months after administration of 30–150 MBq ¹⁷⁷Lu-octreotate. * Statistically significant regulation (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0136204#sec005" target="_blank">Materials and Methods</a>)</p

Blood urea nitrogen (BUN) does not differ in untreated adriamycin mice compared with MC1R agonist treated mice.

<p>BUN was measured in plasma samples taken from mice 8–10 days after injection of adriamycin. There was no difference between the groups (n = 9–10, n.s.). Results are presented as mean ± SEM.</p

Radiation responsive genes.

<p>* Previously proposed kidney damage marker</p><p>^† Previously proposed kidney damage marker and ionizing radiation marker</p><p>^‡ Previously proposed ionizing radiation marker.</p><p><b>Bold</b> indicates that the gene has also been proposed at the protein level to be a potential biodosimeter</p><p>Differentially regulated transcripts which have previously been proposed as biomarkers for kidney injury, radiation induced damage, and radiation biodosimetry [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0136204#pone.0136204.ref033" target="_blank">33</a>–<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0136204#pone.0136204.ref036" target="_blank">36</a>]. The log₂ratio is shown for each gene.</p