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Real-time quantitative PCR analysis of lungs from Litomosoides sigmodonis-infected mice

ABSTRACT: Real-time quantitative PCR analysis of lungs from Litomosoides sigmodonis-infected mice

PROVIDER: PRJNA475552 | ENA |

REPOSITORIES: ENA

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Real-time quantitative PCR analysis of lungs from Litomosoides sigmodonis-infected mice

Project description:A pool of cDNA from naive BALB/c mice lungs (n=8) was compared with one from D70 p.i. Litomosoides sigmodontis-infected mice (n=8) by profiling 84 cytokine-related genes simultaneously. Screening of inflammatory lung environment was performed with a qRT-PCR array (Mouse Cytokines & Chemokines RT2 Profiler PCR Array, Qiagen, Germany) according to manufacturer’s instructions.

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Project description:Balb/cJ mouse received control RNAi or RNAi to 8-oxoguanine DNA glycosylase (Ogg1) intranasally prior to the exposure for 1 h to a) glucose oxidase (1 mU) to induce OGG1-BER or b) OGG1-BER product 8-oxoguanine (1 M-NM-<M). We used SABiosciences Mouse Inflammatory Cytokines & Receptors PCR Array (PAMM-011A) to quantitate inflammatory gene expression dependent on OGG1 and its product 8-oxoguanine. After intranasal challenge with glucose oxidase or 8-oxoguanine, mice were sacrificed and lungs were collected and processed to obtain RNA. Total pooled (n=5) RNA (1 M-NM-<g) was reverse transcribed into cDNA using SuperscriptM-BM-. III First Strand Synthesis System (Invitrogen), mixed with equal amounts of 2X SYBR Green Supermix (Qiagen) and 20 M-NM-<l of reaction mixture was added to each well of the PAM-011A array. The reaction was evaluated using an ABI PRISMM-BM-. 7000 Sequence Detection System using recommended settings by SABiosciences.

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