Il4 mm00445259

Total RNA was isolated from liver tissue (RNeasy Plus Mini Kit, Qiagen) and reversely transcribed into cDNA using a High-capacity cDNA Reverse Transcriptase Kit (Thermo Fisher) according to the manufacturer's instructions. Real-time PCR (RT-PCR) was performed using the following TaqMan Gene Expression Assays: Col1a2 Mm00483888; Acta2 Mm00725412; Mmp2 Mm00439498; Timp1 Mm01341361; Il13 Mm00434204; Il4 Mm00445259; Infg Mm01168134; and Il10 Mm01288386 (Thermo Fisher). Gene expression values were normalized to the endogenous reference gene Gapdh (Rodent GAPDH control reagent, ThermoFisher) and presented as normalized, relative expression values to naive controls.

Total RNA was isolated from snap frozen liver samples (RNeasy Plus Mini Kit, Qiagen, Germany). The quantity of RNA was measured on Colibri Microvolume Spectrometer (Titertek-Berthold, Germany). 500 ng of total RNA was used to be reversely transcribed into cDNA using High-Capacity cDNA Reverse Transcriptase Kit (ThermoFisher, Germany) according to the manufacturer’s instructions. Each qRT-PCR reaction was performed using 2 µl of cDNA in a final volume of 10 µl. All samples were run in duplicates. RT-PCR was performed using the following TaqMan Gene Expression Assays: il-1β Mm00434228, tnf-α Mm00443258, ifn-γ Mm01168134, il-12a Mm00434169, il-12b Mm00434174_m1, il-4 Mm00445259, acta-2 Mm00725412, il-13 Mm00434204, il-10 Mm01288386 (ThermoFisher, Germany). Cycling was performed using QuantStudio 3 (Thermo Fisher Scientific, Germany) under the following reaction conditions: 50°C for 2 min followed by 95°C for 10 min, 45 cycles at 95°C for 15 s, and at 60°C for 1 min. The ΔΔCt method was utilized for relative quantification (16 (link)). Gene expression values were normalized to the endogenous reference gene GAPDH (Rodent GAPDH control reagent, ThermoFisher, Germany) and presented as normalized expression values relative to naive controls.