Constitutional Nephrin Deficiency in Conditionally Immortalized Human Podocytes Induced Epithelial-Mesenchymal Transition, Supported by -Catenin/NF-kappa B Activation: A Consequence of Cell Junction Impairment?
Analysis of additional EMT marker expressions in wild type human podocytes, upon TGF-1 treatment. The cells were treated as indicated in Figure 1. (a) The rabbit anti-P-cadherin (sc-7893), (b) N-cadherin (sc-7939), (d) anti-Slug (sc-15321) and polyclonal antibodies were purchased from Santa Cruz Biotechnologies (Santa Cruz, CA, USA). (c) The rabbit anti-LOXL2 (A01) was purchased from Abnova (Taiwan). To quantify the protein bands, we scanned the blots and performed a densitometry analysis, using the software NIH ImageJ v. 6.4 (freeware, NIH, MD, USA). (e) Confocal laser scanned immunofluorescence of Snail-1 and Fsp-1 expressions. The cells were treated as indicated and prepared for immunofluorescence analysis as described in Section 2. The goat anti-Snail-1 (sc-10433) and anti-Fsp1 (also known as Mts-1) (sc-19949) antibodies were purchased from Santa Cruz Biotechnologies, (Santa Cruz, CA, USA). The secondary antibodies were conjugated with Alexa Fluor 488 dye (Molecular Probes, Eugene, OR, USA). Nuclei were previously stained with propidium iodide. The figure shows a typical experiment of at least three performed under the same conditions.
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