GFP+ cells were sorted by FACS from COL-Cre;WT, COL-Cre;Tgfbr2fl/fl, TNC-Cre;WT, and TNC-Cre;Tgfbr2fl/fl mice at 3 days post UUO. The primary cells were grown in DMEM/F12 media supplemented with 10% FBS and penicillin/streptomycin antibiotics. At passage 3–4, cells were put in 1%FBS ± various concentrations of TGF-β1 (R&D) for 48 hours, and lysates were made using RIPA buffer plus protease and phosphatase inhibitors (Sigma cocktail). Cells were immortalized by sv40-adenovirus, stimulated with PDGF-BB (R&D Systems), CTGF (gift from Pampee Young), or EGF (Millipore), and imaged with a Nikon Eclipse TE300 inverted scope.
Eclipse te300 inverted scope
Manufactured by Nikon
The Eclipse TE300 is an inverted microscope designed for advanced live-cell imaging. It features a stable and vibration-resistant frame, and supports a range of contrast-enhancing techniques such as phase contrast, fluorescence, and differential interference contrast (DIC). The Eclipse TE300 is suitable for a variety of biological and cell culture applications.
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Lab products found in correlation
2 protocols using eclipse te300 inverted scope
1
Isolation and Characterization of Renal Fibroblasts
2
Isolation and Characterization of Renal Fibroblasts
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GFP+ cells were sorted by FACS from COL-Cre;WT, COL-Cre;Tgfbr2fl/fl, TNC-Cre;WT, and TNC-Cre;Tgfbr2fl/fl mice at 3 days post UUO. The primary cells were grown in DMEM/F12 media supplemented with 10% FBS and penicillin/streptomycin antibiotics. At passage 3–4, cells were put in 1%FBS ± various concentrations of TGF-β1 (R&D) for 48 hours, and lysates were made using RIPA buffer plus protease and phosphatase inhibitors (Sigma cocktail). Cells were immortalized by sv40-adenovirus, stimulated with PDGF-BB (R&D Systems), CTGF (gift from Pampee Young), or EGF (Millipore), and imaged with a Nikon Eclipse TE300 inverted scope.
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