No UCP1 in the kidney

Figures

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  Figure 1. Immunoreactivity to the UCP1 antibody C10 in the perirenal brown adipose tissue (perirenal BAT) and the kidney of mice acclimated to 4 °C. (A–C) Representative confocal images obtained using the rabbit polyclonal antibody raised against the C-terminal decapeptide of UCP1 (C10). (A) Representative confocal image of the transversal section of the kidney, also showing adjacent perirenal BAT. UCP1 – green; perilipin (an adipocyte identity marker) – red; nuclei – blue. Magnified insets provide better visualization of perirenal BAT (inset 1) and the kidney parenchyma (inset 2). Scale bar, 100 μm. (B) and (C) Representative confocal images of the transversal section of the kidney acquired with higher magnification. UCP1 – green; nuclei – blue; note the absence of immunoreactivity for perilipin. Scale bar 50 μm (B) and 10 μm (C). (For interpretation of the references to color/colour in this figure legend, the reader is referred to the Web version of this article.)

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  Figure 2. Immunoreactivity to the UCP1 antibody in the kidney of UCP1-deficient mice suggests antibody cross-reactivity. (A–B) Representative confocal images of the transversal section of the kidney, also showing adjacent perirenal BAT, obtained using the C10 UCP1 antibody. Wild-type (A) and UCP1–KO (B) mice, acclimated to room temperature and approximately two months old, were analysed. UCP1 – green; perilipin (an adipocyte identity marker) – red; nuclei – blue. Magnified insets provide better visualization of perirenal BAT (insets 1 and 3) and the kidney parenchyma (insets 2 and 4). Scale bars, 100 μm. (For interpretation of the references to color/colour in this figure legend, the reader is referred to the Web version of this article.)

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  Figure 3. Immunoreactivity to the commercial UCP1 antibody ab10983 in the kidney of UCP1-deficient mice further suggests that the staining is not due to UCP1. (A–B) Representative confocal images of the transversal section of the kidney, also showing adjacent perirenal BAT, obtained using the rabbit polyclonal antibody from Abcam (ab10983). Wild-type (A) and UCP1–KO (B) mice, acclimated to room temperature and approximately two months old, were analysed. UCP1 – green; nuclei – blue. Magnified insets provide better visualization of the kidney papilla (insets 1 and 3) and the kidney parenchyma (insets 2 and 4). Scale bars, 100 μm. (For interpretation of the references to color/colour in this figure legend, the reader is referred to the Web version of this article.)

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  Figure 4. UCP1 is not expressed in the kidney. (A) UCP1 mRNA levels in IBAT (interscapular BAT), ingWAT (inguinal WAT), gWAT (gonadal WAT), liver and kidney of wild-type (n = 3) and UCP1–KO (n = 3) mice acclimated to room temperature and approximately two months old. Each symbol represents a sample from one mouse. Values are antilog-transformed Cq values (2−Cq) presented as means ± SEM. For each tissue, the mean Cq value is reported. Higher Cq values thus indicate lower expression levels. (B–C) Representative confocal images of the transversal section of the kidney, also showing adjacent perirenal BAT, obtained using the rabbit monoclonal antibody (E9Z2V) from Cell Signaling Technology (72298S). Wild-type (B) and UCP1–KO (C) male mice, acclimated to room temperature and approximately two months old, were analysed. UCP1 – green; perilipin (an adipocyte identity marker) – red; nuclei – blue. Note the complete absence of immunoreactivity to this UCP1 antibody in both the kidney and the perirenal BAT of UCP1-deficient mice (C). Magnified insets provide better visualization of perirenal BAT (insets 1 and 3) and the kidney parenchyma (insets 2 and 4). Scale bars, 100 μm. (For interpretation of…