Description
To identify novel transcriptional targets of beta-catenin and FoxO1 and FoxO3 in renal epithelial cells, we used conditionally immortalized murine proximal tubule (PT) cells (these cells, from Tgfbr2floxed mice, are described in detail in manuscript PMID: 23160515 in which Leslie Gewin is first author, JASN 2012). PT cells were either treated with Wnt3a (to activate beta-catenin) or the control diluting buffer, H2O2 to induce oxidative stress, and some were transfected with FoxO1, FoxO3, FoxO1 and 3, or scramble siRNA prior to Wnt and H2O2 treatment.
Overall Design
There were a total of 10 samples, replicates were not done. For publication, we analyzed sample 7 compared to 8 and 7 compared to 9. In other words, we compared the transcripts of PT cells with FoxO1 siRNA plus H2O2 and Wnt3a to PT cells with scramble siRNA and H2O2 and Wnt3a. Also, we compared transcripts of PT cells with FoxO3 siRNA plus H2O2 and Wnt3a to PT cells with scramble siRNA and H2O2 and Wnt3a. Separately, we also did 2 other analyses: 1) compare the genes upregulated by Wnt in regular conditions (PT media) versus oxidative stress (H2O2). This would be comparing the genes that change from sample 1 to 2 and comparing with the genes that change from sample 5 to 6. 2) We also compared the effect of knocking down FoxO1/3 in PT media with those in oxidative stress (comparing the change from sample 3 to 4 with that of sample 7 to 10).; Raw data could not be provided for this submission because the files had been previously deleted
Curator
xm_li