Journal: Physiological Reports
Article Title: Renal stromal miRNAs are required for normal nephrogenesis and glomerular mesangial survival
doi: 10.14814/phy2.12537
Figure Lengend Snippet: Increased apoptosis in FoxD1 GC ; Dicer fl/fl stromal derived cells. (A) ToppFun analysis of differentially expressed transcripts obtained from E18.5 control and FoxD1 GC ; Dicer fl/fl kidneys by microarray showed evidence of increased expression of transcripts associated with apoptosis and dysregulation of the p53 signaling pathway. (B) Quantitative PCR validated the upregulation of Bcl2 l11 (Bim) and genes associated with p53 signaling including Trp53inp1, Bax, Jun, Cdkn1a (p21), Mmp2, and Arid3a in E18.5 FoxD1 GC ; Dicer fl/fl kidneys. n = 6, * P < 0.05, ** P < 0.01, *** P < 0.001. (C) Western blot analysis showed increased levels of BimEL and BimL but normal levels of p53 in E18.5 FoxD1 GC ; Dicer fl/fl kidneys. (D, E) Section immunohistochemistry for confirmed increased and ectopic Bim expression (brown) in the renal stroma and nephron progenitors. (F–K) Section in situ hybridization revealed increased and ubiquitous expression of Trp53inp1, Bax, and p21 in E18.5 FoxD1 GC ; Dicer fl/fl kidneys. (L, M) Immunofluorescence revealed increased aCaspase3 + (green) apoptotic cells in the E18.5 FoxD1 GC ; Dicer fl/fl kidneys. (N–Q) Apoptotic cells were largely localized to Meis1,2,3 + (red) glomerular mesangium and interstitial cells. (R, S) No differences in proliferation based on phosphohistone H3 staining were observed. Immunostaining was performed on at least three control and mutant embryos.
Article Snippet: Whole-transcriptome microarray profiling was performed using an Illumina MouseWG-6 v2.0 chip on three litters of E15.5 and E18.5 kidneys.
Techniques: Derivative Assay, Control, Microarray, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Immunohistochemistry, In Situ Hybridization, Immunofluorescence, Staining, Immunostaining, Mutagenesis