rabbit anti ter 119  (R&D Systems)

Journal: The FASEB Journal

Article Title: Loss of Nfat5 promotes lipid accumulation in vascular smooth muscle cells

doi: 10.1096/fj.202100682r

Figure Lengend Snippet: FIGURE 3 Analysis of the responses of mouse aortic VSMCs to cholesterol stimulation. VSMCs isolated from aortae of Nfat5fl/fl mice were exposed to cholesterol-supplemented medium (Panserin, chol:MbCD 10 µg/mL) or the respective solvent control for three days. Abca1, Acat1, Hmgcr, Lgals3, and Cd68 expression was analyzed by real-time quantitative PCR (A). The expression of the housekeeping gene Rps12 served as internal control (A, *P < .05, ***P < .001 vs control, n = 6-7). NFAT5 protein was detected by capillary electrophoresis in cytosolic and nuclear fractions of cell lysates B and C. HDAC1 and α-tubulin served as loading controls for the nuclear and cytosolic fractions respectively (C, Cytosol: n.s. vs control; Nucleus: *P < .05 vs control, n = 3, Ctr., control; Chol., cholesterol; n.s., not significant)

Article Snippet: Oil Red O (ORO)- stained regions from the aortae of N5fl/fl and N5(SMC)−/− mice fed an atherogenic diet for 25 weeks and appropriate regions from aortae of normal chow dietfed N5fl/fl and N5(SMC)−/− mice (see ORO staining) were Target Host Dilution Cat. no.; supplier Application Primary antibodies Anti- alpha- Tubulin Rabbit 1:10 2144; Cell Signaling Technology Wes Anti- HDAC1 Rabbit 1:10 NB100- 56340; Novus Biologicals Wes Anti- histone H3 Rabbit 1:1000 ab1791; Abcam Wes Anti- NFAT5 Mouse 1:10 SC- 398171; Santa Cruz Wes Anti- NFAT5 Rabbit 1:100 NB120- 3446; Novus Biologicals IF Anti- NFAT5 Rabbit 1:500 NB120- 3446; Novus Biologicals Wes Anti- VCP Mouse 1:50 ab11433; Abcam Wes Secondary antibodies Anti- Mouse HRP Goat – 042- 205; Bio- Techne Wes Anti- Rabbit IgG- Cy3 Donkey 1:100 711- 166- 152; Dianova IF Abbreviations: IF, immunofluorescence; Wes, capillary electrophoresis.

Techniques: Isolation, Solvent, Control, Expressing, Real-time Polymerase Chain Reaction, Electrophoresis