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MB Sample ID: SA172714
| Local Sample ID: | PLKO_3_Hilic |
| Subject ID: | SU001927 |
| Subject Type: | Cultured cells |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
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Collection:
| Collection ID: | CO001920 |
| Collection Summary: | Briefly, podocytes were cultured on BD BioCoat Collagen I plates (BD Biosciences, San Jose, CA) at 33°C in RPMI 1640 complete media with 20 U/ml mouse recombinant IFN-g (Thermo Fisher, Carlsbad, CA). To induce differentiation, we cultured podocytes in DMEM (5.5mM glucose and 5% FBS) at 37°C without IFN-g for 10-12 days. To rescue the expression of PGC1-a in stable Tug1-knockdown CRISPR clone (Tug1-KD)(Long et al., 2016), CMV enhancer/promoter-driven mouse Pgc1-a cDNA (Addgene, Watertown, MA) was inserted into vector Zeo-pT-MCS-GFP-T2A-Puro(Long et al., 2020), a modified PiggyBac transposon system, selected with 1ug/ml puromycin and sorted by GFP, to generate Tug1-KD/Pgc1-OE. We isolated kidney podocytes by positive selection with biotin-labelled Kirrel3 and Podocalyxin antibodies (2.5 μg/antibody/mouse, R&D Systems, Minneapolis, MN) followed by Streptavidin M-280 Dynabeads as previously described (Badal et al., 2016). |
| Sample Type: | Epithelial cells |
