Journal: Journal of Lipid Research

Article Title: Deficiency in ZMPSTE24 and resulting farnesyl–prelamin A accumulation only modestly affect mouse adipose tissue stores [S]

doi: 10.1194/jlr.RA119000593

Figure Lengend Snippet: Prelamin A accumulates in adipocytes of LmnaPLAO/PLAOZmpste24fl/flAdipoq-Cre+ mice, as judged by immunofluorescence microscopy. Tissue sections of gonadal WAT, BAT, and liver from a LmnaPLAO/PLAOZmpste24fl/flAdipoq-Cre– mouse and a LmnaPLAO/PLAOZmpste24fl/flAdipoq-Cre+ mouse were stained with antibodies against prelamin A (green), lamin A/C (red), and collagen IV (cyan). DNA was stained with DAPI (blue). Liver from a Zmpste24−/− mouse was included as a positive control for the prelamin A antibody. Images were recorded with a confocal microscope using a 20× objective and a 2× digital zoom. Identical microscope settings were used for all three tissue samples. Scale bar, 20 μm.

Article Snippet: Primary antibodies included a rat anti-prelamin A monoclonal antibody (clone 3C8; 12 μg/ml) ( 18 ), a goat anti-CD31 antibody (Novus, AF3628; 4 μg/ml), a rabbit anti-collagen IV antibody (Abcam, ab19808; 1:300), and a mouse anti-lamin A/C antibody (Santa Cruz Biotechnology, sc-375248; 1:500) labeled with Alexa 647 (Thermo Fisher).

Techniques: Immunofluorescence, Microscopy, Staining, Positive Control

Journal: Journal of Lipid Research

Article Title: Deficiency in ZMPSTE24 and resulting farnesyl–prelamin A accumulation only modestly affect mouse adipose tissue stores [S]

doi: 10.1194/jlr.RA119000593

Figure Lengend Snippet: Levels of prelamin A protein and Lmna gene expression in WAT are higher in male mice than female mice. A: Western blots on tissue extracts from female and male LmnaPLAO/PLAOZmpste24fl/flAdipoq-Cre+ mice. Tissue extracts from a Zmpste24+/+ (WT) and Zmpste24−/− (ZmpKO) mice were included as controls. Prelamin A was detected with a prelamin A–specific antibody (green, top). Lamin A, lamin C, and prelamin A were detected with a lamin A/C antibody (red, middle). Tubulin was measured as a loading control (green, bottom). B: Quantification of prelamin A expression, normalized to tubulin, in the Western blots shown in panel A. Graph shows the mean ± SEM; n = 3 mice per group). *P < 0.02. C, D: Quantitative RT-PCR studies of total Lmna expression (prelamin A and lamin C) and prelamin A expression in gonadal WAT (C) and liver (D) tissue from female (gray) and male (red) LmnaPLAO/PLAOZmpste24fl/flAdipoq-Cre+ and LmnaPLAO/PLAOZmpste24fl/flAdipoq-Cre– mice. Data were normalized to cyclophilin A. Data show the mean ± SEM for 9 female mice and 12 male mice. *P < 0.05.

Article Snippet: Primary antibodies included a rat anti-prelamin A monoclonal antibody (clone 3C8; 12 μg/ml) ( 18 ), a goat anti-CD31 antibody (Novus, AF3628; 4 μg/ml), a rabbit anti-collagen IV antibody (Abcam, ab19808; 1:300), and a mouse anti-lamin A/C antibody (Santa Cruz Biotechnology, sc-375248; 1:500) labeled with Alexa 647 (Thermo Fisher).

Techniques: Gene Expression, Western Blot, Control, Expressing, Quantitative RT-PCR