pone.0229397.g001.tif (511.02 kB)

Generation of podocyte-specific Tsc2 knockout mice, Tsc2Δpodocyte. Homozygous floxed Tsc2 mice were crossed with Nphs2-Cre transgenic mice to generate Tsc2Δpodocyte, Nphs2-Cre+/-; Tsc2flox/flox.

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posted on 19.03.2020, 17:41 by Wakiko Iwata, Hiroyuki Unoki-Kubota, Hideki Kato, Akira Shimizu, Michihiro Matsumoto, Toshiyuki Imasawa, Arisa Igarashi, Kenji Matsumoto, Tetsuo Noda, Yasuo Terauchi, Masaomi Nangaku, Masato Kasuga, Yasushi Kaburagi

(A) PCR genotyping of genomic DNA from mouse tails. NPHS2-Cre+/-, Tsc2wt/wt mice (Nphs2-Cre) and NPHS2-Cre-/-, Tsc2flox/flox mice (Tsc2flox/flox) were used as controls. F, flox; +, wt. (B) Primary cultured podocytes isolated from Tsc2Δpodocyte and wild-type control mice at 4 weeks of age demonstrated that Tsc2 mRNA was knocked down by over 80% in the podocytes of Tsc2Δpodocyte mice compared with control mice. Results are expressed as means ± s.d. *P < 0.05 compared with age-matched controls. (C) Western blot analysis of primary cultured podocytes isolated from Tsc2Δpodocyte and control mice. TSC2 gives a band at 200 kDa (arrow). Non-specific bands were also detected above the TSC2 band in all samples. β-Tubulin was used as an internal control. (D) Tsc2 mRNA expression in various tissues of Tsc2Δpodocyte and wild-type controls. Expression levels of Tsc2 mRNA in Tsc2Δpodocyte were comparable with those in controls in all the tissues examined, including the renal cortex. White bar, Nphs2-Cre; dotted bar, Tsc2flox/flox; black bar, Tsc2Δpodocyte.