Journal: Journal of Cellular and Molecular Medicine
Article Title: The transcription factor Dach1 is essential for podocyte function, et al. The transcription factor Dach1 is essential for podocyte function
Figure Lengend Snippet: Effect of Dach1 expression in PEC s on synaptopodin and further PEC /podocyte proteins. A, The up‐regulation of synaptopodin (red) and co‐localization with F‐actin (green) was shown by immunofluorescence and super resolution microscopy in PEC ‐Dach1. B, Synaptopodin‐positive cells are significantly increased in PEC ‐Dach1 as compared to PEC ‐Ctrl (upper diagram, n = 5 experiments with > 100 cells per experiment). Synaptopodin‐positive PEC s were almost always positive for Dach1 expression (lower diagram, n = 5 experiments with > 100 cells). C, Podocyte marker genes ( Wt1 , Nphs2 ), PEC marker genes ( Pax2 , Cav1 ) and differentiation/development regulators ( Eya1, Eya3 ) were quantified by qRT ‐ PCR analysis of total mRNA isolated from PEC ‐Ctrl and PEC ‐Dach1. qRT ‐ PCR experiments were normalized to PEC ‐Ctrl (dashed line). Actin , Gapdh and 18s RNA served as reference genes (n ≥ 3, means ± SEM ). D, WT 1 up‐regulation and Pax2 down‐regulation in PEC ‐Dach1 were shown by Western blot analysis. E, Relative Wt1 and Pax2 protein levels on Western blots were quantified for PEC ‐Ctrl (white bars) and PEC ‐Dach1 (black bars) and revealed an up‐regulation of WT 1 (n = 4, means ± SEM ) and a down‐regulation of Pax2 (n = 6, means ± SEM ) in PEC ‐Dach1. Western blots were normalized to PEC ‐Ctrl and Gapdh levels. *** P
Article Snippet: The quantitative real‐time PCR (qRT‐PCR) analysis was performed on a LightCycler Nano (Roche Diagnostics GmbH, Mannheim, Germany) using the iTaq Universal SYBR Green Supermix (Bio‐Rad, Hercules, CA, USA) with Actb and Gapdh as reference genes.
Techniques: Expressing, Immunofluorescence, Microscopy, Marker, Quantitative RT-PCR, Isolation, Western Blot