Genome-wide analysis of gene expression in normal and fibrotic mouse kidneys with and without Twist1 or with and without Snai1(Snail) expression in proximal tubular epithelial cells

In this study mice were engineered to specifically delete Twist1 or Snail expression in proximal tubular epithelial cells of the kidney (ggt-cre+;Twist flox/flox and ggt-cre+;Snail flox/flox ). These mice and control mice (ggtcre-;Twist flox/flox: these express Twist1, and ggtcre-;Snail flox/flox:these express Snail) were subjected to unilaterial ureteric obstruction. This experiment allows for the collection and analysis of expression in contralateral healthy (HK) kidney and obstructed disease (DK) kidney. Total RNA was isolated from the contralateral healthy (HK) and obstructed disease (DK) kidneys of 3 mice with ggt-cre-;Twist flox/flox genotype (Wildtype like) and 4 mice with ggt-cre+;Twist flox/flox genotype (loss of Twist1 in proximal tubular epithelial cells), 3 mice with ggt-cre-;Snail flox/flox genotype (Wildtype like) and 3 mice with ggt-cre+;Snail flox/flox genotype (loss of Snail in proximal tubular epithelial cells). Total RNA was also isolated from kidneys of completely healthy mice: 3 mice ggt-cre-;WT, 3 mice ggt-cre+;WT, 3 mice ggt-cre+;Twist flox/flox and 3 ggt-cre+;Snail flox/flox本研究中，我们构建了工程化小鼠模型，可特异性敲除肾脏近端肾小管上皮细胞（proximal tubular epithelial cells）内Twist1或Snail的基因表达，具体品系为ggt-cre+;Twist flox/flox与ggt-cre+;Snail flox/flox。随后将上述工程化小鼠与对照小鼠均接受单侧输尿管梗阻（unilateral ureteric obstruction）造模处理，其中对照小鼠分为两组：ggt-cre-;Twist flox/flox（该组可正常表达Twist1）与ggt-cre-;Snail flox/flox（该组可正常表达Snail）。本实验可收集对侧健康肾脏（contralateral healthy kidney，HK）与梗阻病变肾脏（obstructed disease kidney，DK）样本，并对其中的基因表达情况进行分析。我们从以下基因型小鼠的HK与DK肾脏中分离总RNA（total RNA）：3只ggt-cre-;Twist flox/flox基因型小鼠（野生型类似组）、4只ggt-cre+;Twist flox/flox基因型小鼠（近端肾小管上皮细胞Twist1敲除组）、3只ggt-cre-;Snail flox/flox基因型小鼠（野生型类似组）与3只ggt-cre+;Snail flox/flox基因型小鼠（近端肾小管上皮细胞Snail敲除组）。此外，我们还从完全健康小鼠的肾脏中分离总RNA，包括3只ggt-cre-;WT小鼠、3只ggt-cre+;WT小鼠、3只ggt-cre+;Twist flox/flox小鼠与3只ggt-cre+;Snail flox/flox小鼠。