Mice were subjected to IMV (peak inspiratory pressure 45, PEEP 3, RR 80, FiO2 100%) or control ventilation (peak inspiratory pressure 12, PEEP 3, RR 80, FiO2 100%) , duration of mechanical ventilation 4 hours. After that alveolar type II epithelial cells were isolated form lungs
Growth protocol
n/a
Extracted molecule
total RNA
Extraction protocol
Total RNA was extracted with RNeasy Mini Kit followed by DNase I treatment.
Label
SYBR Green
Label protocol
PCR were performed with Qiagen RT2 prolfer assay according to the manufacturer's instruction
Hybridization protocol
n/a
Scan protocol
n/a
Description
Ctrl
Data processing
The normalization and all the data analysis were performed according to the manufacturers instructions using their web-based software package: http://sabiosciences.com/pcrarraydataanalysis.php For the normalization it uses the average of four housekeeping genes: B2M, Gusb, ACTB, Hsp90ab1 Target gene signals normalized to housekeeping genes; 2^-deltaCt, where deltaCt = (Ct_Target ? Ct_HKG)] The web-based software package automatically performs all deltadeltaCt based fold-change calculations from the uploaded raw threshold cycle data. Matrix normalized worksheet reports normalized signal (against housekeeping genes). Fold Change worksheet reports test/control (i.e., Ctrl/IMV) ratios.
Changes in carbohydrate metabolism in alveolar epithelial type II cells in Ub-Cre and Hif1a loxp/loxp Ub cre mice in respnse to injurious mechanical ventilation (IMV)
