Gene Expression Analysis of huPCLS

Total RNA was isolated from huPCLS and qPCR analysis was performed as previously described [15 (link),25 (link),36 (link)] using individual TaqMan^® Probe-Based Gene Expression Assays (Applied Biosystems, Life Technologies, Carlsbad, CA, USA) selected for proinflammatory cytokines (IL-1β, IL-6, TNFα, and IL-1α), inflammation (COX-2), relevant cytoprotective and antioxidant enzymes (heme oxygenase-1 (HMOX1) and NADPH:quinone oxidoreductase-1 (NQO1), and cell cycle markers (TP53, CDK2, CDKN1A, CDKN2A, CDK4, CDK6, RB, and E2F). Transcript levels of tested genes were normalized to β-Actin.

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Velalopoulou A., Chatterjee S., Pietrofesa R.A., Koziol-White C., Panettieri R.A., Lin L., Tuttle S., Berman A., Koumenis C, & Christofidou-Solomidou M. (2017). Synthetic Secoisolariciresinol Diglucoside (LGM2605) Protects Human Lung in an Ex Vivo Model of Proton Radiation Damage. International Journal of Molecular Sciences, 18(12), 2525.

Publication 2017

TaqMan® Probe-Based Gene Expression Assays

Actin Antioxidant Assays Cdk2 Cdk6 Cdkn1a Cdkn2a Cell cycle Cox 2 Cytokines Enzymes Gene expression Genes Heme oxygenase 1 Il 1β Inflammation Nadph Quinone oxidoreductase 1 Tnfα Tp53

Corresponding Organization : University of Pennsylvania

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Variable analysis

independent variables

Not explicitly mentioned

dependent variables

Transcript levels of proinflammatory cytokines (IL-1β, IL-6, TNFα, and IL-1α)
Transcript levels of inflammation-related genes (COX-2)
Transcript levels of cytoprotective and antioxidant enzymes (HMOX1 and NQO1)
Transcript levels of cell cycle markers (TP53, CDK2, CDKN1A, CDKN2A, CDK4, CDK6, RB, and E2F)

control variables

Not explicitly mentioned

controls

Transcript levels were normalized to β-Actin, which may serve as a reference or endogenous control

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