Skip to main content
Figure 3 | Molecular Cancer

Figure 3

From: USP1 deubiquitinase: cellular functions, regulatory mechanisms and emerging potential as target in cancer therapy

Figure 3

USP1 gene mutations and altered USP1 mRNA expression in lung cancer. A. Shematic representation of USP1 protein showing the position of 13 lung cancer-associated USP1 mutations reported to date (5 April 2013) in the COSMIC mutation database. Amino acid changes are indicated using the one-letter code. B. Relative USP1 mRNA expression normalized to GAPDH in paired samples of tumor tissue and adjacent normal tissue from four NSCLC patients (represented by different symbols), showing higher USP1 expression in tumor tissue. Total RNA was isolated using Trizol (Invitrogen), and complementary DNA (cDNA) was synthesized using the Dynamo cDNA synthesis kit for qRT-PCR (Thermo Scientific). Quantitative real-time PCR (qRT-PCR) was performed using the TaqMan Universal Master Mix (Applied Biosystems) according to the manufacturer’s instructions on an ABI Prism 7500 instrument. Assay-on-Demand Gene expression primers and probes (Applied Biosystems) were used to specifically amplify USP1 (Hs00163427_m1) and Human GAPD (GAPDH) Endogenous Control (4326317E). C. Graph comparing the relative expression of USP1 mRNA normalized to GAPDH internal control in normal lung tissue (white bar; four individual samples) with the expression in pooled samples of the three different NSCLC histological subtypes (grey bar; adenocarcinoma, squamous cell carcinoma and large cell carcinoma). Each pooled sample included laser-microdissected specimens from five different patients, as described previously [88]. qRT-PCR analysis was carried out as described above. Graph bars show mean ± SEM. USP1 was significantly overexpressed in the pooled tumor samples (***P < 0.001). D. Relative USP1 mRNA expression normalized to GAPDH in 20 different NSCLC cell lines compared to the average of four normal lung tissue mRNA samples. qRT-PCR analysis was carried out as described above. The relative mRNA levels were determined with the ΔΔCt method using the “normal” value as a reference. Graph bars show mean ± SD of two replicates.

Back to article page