Taqman gene expression assays specific pcr primers sets

Total RNA was extracted from murine lung tissues using the Qiagen RNeasy Plus mini kit (Qiagen, Hilden, Germany) according to the supplier's instructions, and further treated with the Qiagen RNase-Free DNase Set (Qiagen) to remove genomic DNA. For the integrity measurement, 4 µl of the total RNA were analyzed on an Agilent Cary 60 UV-VisSpectrophotometer (Agilent Technologies Inc., Santa Clara, CA). Total RNA was transcribed into cDNA and amplified using the Express One-Step Superscript qRT-PCR Kit (Invitrogen, Carlsbad, CA). Analysis of murine CXCL1 (KC), CCL20 and Mideline-1 (MID-1) was carried out with validated TaqMan Gene Expression Assays specific PCR primers sets (Invitrogen) [26] (link). All samples were run in triplicate using the real time thermal analyzer Rotor-Gene 6000 (Corbett, Cambridge, UK). Expression values were normalized to the housekeeping murine gene POLR2A amplified in the same sample.

Total RNA was extracted from cells using the QIAGEN RNeasy Plus mini kit (QIAGEN, Hilden, Germany), accordingly to the supplier's instructions. Total RNA was transcribed into cDNA and amplified using the Express One-Step Superscript qRT-PCR Kit (Invitrogen, Carlsbad, CA). Analysis of human CDKN1A, MDM2 and BAX gene expression was carried out with validated TaqMan Gene Expression Assays specific PCR primers sets (Invitrogen). All samples were run in triplicate using the real time thermal analyzer Rotor-Gene™ 6000 (Corbett, Cambridge, UK), as previously described [47 (link)]. Expression values were normalized to the housekeeping gene POLR2A amplified in the same sample.