RNA Isolation and Gene Expression Analysis

Total RNA from tumor cell lines or tissues was isolated with the RNeasy Mini Kit (Qiagen, Germantown, MD, USA), according to the manufacturer’s instructions [18 (link)]. cDNA preparation was conducted according to standard procedures, using M-MLV Reverse Transcriptase (Promega, Madison, WI, USA) and oligo-dT primers (Promega, Madison, WI, USA). Gene expression was measured using the following Taqman gene-specific probes from Thermo Fisher Scientific: NRP-1 (Hs00826128_m1), EGFR (Hs00193306m1), and the housekeepers GAPDH (Hs04420632_g1), and

β

-actin (Hs99999903_m1) [19 (link)].

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Napolitano V., Russo D., Morra F., Merolla F., Varricchio S., Ilardi G., Di Crescenzo R.M., Martino F., Mascolo M., Celetti A., Tamagnone L, & Staibano S. (2021). Neuropilin-1 Expression Associates with Poor Prognosis in HNSCC and Elicits EGFR Activation upon CDDP-Induced Cytotoxic Stress. Cancers, 13(15), 3822.

Publication 2021

RNeasy Mini Kit M-MLV Reverse Transcriptase oligo-dT primers Taqman gene-specific probes

Actin Cdna Egfr Gapdh Gene Gene expression Oligo dt Primers Promega Reverse transcriptase Tissues Tumor cell lines

Corresponding Organization : University of Molise

Other organizations : Università Cattolica del Sacro Cuore, University of Naples Federico II, Institute for Experimental Endocrinology and Oncology, Istituti di Ricovero e Cura a Carattere Scientifico, Agostino Gemelli University Polyclinic

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Variable analysis

dependent variables

Gene expression of NRP-1
Gene expression of EGFR

control variables

GAPDH
β-actin

controls

Positive control: N/A
Negative control: N/A

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