kidney distal convoluted cells  (ATCC)

Journal: American Journal of Nuclear Medicine and Molecular Imaging

Article Title: Development of zirconium-89 PET for in vivo imaging of alpha-klotho

doi:

Figure Lengend Snippet: Autoradiograph of distal convoluted tubule kidney cell line featuring total binding and non-specific binding. A. Total binding of [89Zr]DFO-anti-mKlotho. B. Non-specific binding measured in the presence of 5 mg/mL of anti-mKlotho. C. The chart summarizes radioactive uptake (digital light units, DLU/mm2) of the [89Zr]DFO-anti-mKlotho with calculated total, non-specific and specific binding values. The specific binding of [89Zr]DFO-anti-mKlotho in the cultured kidney cell line, derived from blocking studies with unlabeled anti-mKlotho antibody, was 30-40%.

Article Snippet: Kidney distal convoluted cells (catalog# ATCC CRL-3250, American Type Culture Collection, Manassas, VA) were expanded and 12 microfuge tubes containing 6.8 million cells each were incubated with 703 MBq [ 89 Zr]DFO-anti-mKlotho per tube.

Techniques: Autoradiography, Binding Assay, Cell Culture, Derivative Assay, Blocking Assay

Journal: Nature Communications

Article Title: Discovery and preclinical evaluation of anti-miR-17 oligonucleotide RGLS4326 for the treatment of polycystic kidney disease

doi: 10.1038/s41467-019-11918-y

Figure Lengend Snippet: RGLS4326 inhibits miR-17 and de-represses direct miR-17 targets. a RGLS4326 (green triangle) dose-responsively inhibits miR-17 in HeLa cell luciferase assay 24 h after transfection, with an EC 50 value of 28.3 ± 4.0 nM (mean ± standard deviation, n = 7 independent experiments). b RGLS4326 dose-responsively de-represses multiple miR-17 target genes (as measured by miR-17 PD-Sig) in mouse IMCD3 cells 24 h after transfection, with an EC 50 value of 77.2 ± 20.2 nM ( n = 4 independent experiments). c – d RGLS4326 treatment results in de-repression of the direct miR-17 target genes, Pkd1 and Pkd2 in IMCD3 cells ( n = 3). e RGLS4326, but not control oligo, functionally inhibited miR-17 and de-repressed miR-17 PD-Sig in six cell lines derived from normal (DBA-WT, M1, MDCT, LTL-WT) or PKD (DBA-PKD and LTL-PKD) mouse kidneys after 24 h treatment by transfection at 30 nM ( n = 3) and in f normal mouse kidney tissue slice culture after 72 h ex vivo incubation at 10 μM ( n = 4). Control oligo (grey triangle) containing the same chemical-modification, length, and design as RGLS4326, but different base pair sequence, was used as a negative control. Error bars represent standard deviations. * p < 0.05, **** p < 0.0001. One-way ANOVA, Dunnette’s multiple comparison test. Source data for Fig. 2 a – f is provided in Source data files

Article Snippet: Mouse MDCT kidney distal convoluted tubule cells (ATCC CRL-3250) were cultured in 1:1 DMEM/F-12 supplemented with 5% FBS, 1 g L −1 glucose, and 1 mM sodium pyruvate.

Techniques: Luciferase, Transfection, Standard Deviation, Control, Derivative Assay, Ex Vivo, Incubation, Modification, Sequencing, Negative Control, Comparison