Journal:

Article Title: Improved Hepatic Gene Transfer by Using an Adeno-Associated Virus Serotype 5 Vector

doi: 10.1128/JVI.76.20.10497-10502.2002

Figure Lengend Snippet: Estimation of vector gene copy number in AAV-transduced livers by quantitative PCR (3 months after vector administration). A 350-bp fragment of the hF.IX cDNA as present in the AAV-EF1α-hF.IX vector was coamplified with a 1.1-kb fragment from the endogenous murine HPRT gene using biotinylated primers (20 cycles of 95°C for 1 min, 56°C for 1 min, and 72°C for 2 min, separated on a 2% agarose gel, transferred to a nylon membrane, and visualized with the Southern light detection system from Applied Biosystems. Template for PCR was as follows: 100 ng of genomic DNA extracted from several random pieces of liver and subsequently combined for each individual animal. Each sample column represents an individual animal. standards, linearized plasmid pAAV-EF1α-hF.IX (0.01, 0.1, or 1 pg) mixed with 100 ng of genomic mouse DNA (extracted from untransduced animal); NC, negative control (template, genomic DNA from untransduced animal). Bands were analyzed by densitometric scanning, and intensities were quantitated with NIH Image 6.16 software. Shown is one representative blot. Ratios of band intensities (hF.IX band/mAAT band) are for the blot shown. Gene copy number estimates are average for two experiments.

Article Snippet: A 350-bp fragment of the hF.IX cDNA as present in the AAV-EF1α-hF.IX vector was coamplified with a 1.1-kb fragment from the endogenous murine HPRT gene using biotinylated primers (20 cycles of 95°C for 1 min, 56°C for 1 min, and 72°C for 2 min, separated on a 2% agarose gel, transferred to a nylon membrane, and visualized with the Southern light detection system from Applied Biosystems.

Techniques: Plasmid Preparation, Real-time Polymerase Chain Reaction, Agarose Gel Electrophoresis, Negative Control, Software

Journal: Cancer Research

Article Title: AREG and EREG Are Predictive Biomarkers of Response to EGFR Inhibition in Gastroesophageal Cancer

doi: 10.1158/0008-5472.CAN-25-0073

Figure Lengend Snippet: A subset of gastroesophageal tumors not displaying EGFR gene amplification is sensitive to anti-EGFR drugs. A, Heatmap showing the viability (in percentage vs. untreated) of 27 primary cell lines treated for 6 days with the indicated doses of the mAb cetuximab. The heatmap represents the average of three independent experiments. B, Boxplot showing the sensitivity of the 27 primary cells (divided into resistant and sensitive groups on the basis of the response to cetuximab) to the indicated drugs. Each dot represents the mean AUC computed by the mean of three independent experiments. Statistical significance was calculated using the Student t test. The labeled dot indicates a HER2-amplified cell line (GTR0233). C, Boxplots showing the percentage of nonviable cells (positive for trypan blue staining), untreated (−) cells, or cells treated (+) with cetuximab (CTX; 5 µg/mL for 6 days). The average of four independent experiments performed on three cetuximab-sensitive (GTR0539, GTR0640, and GTR0247) and three cetuximab-resistant cell lines (GTR0125, GTR0498, and GTR0687) is shown. Statistical significance was calculated using the ratio paired t test. *, P < 0.05; **, P < 0.01; ****, P < 0.0001.

Article Snippet: The following primary antibodies were used: HER3 (Cell Signaling Technology #12708 RRID: AB_2721919), HER2 (Cell Signaling Technology #2165 RRID: AB_10692490), vinculin (Millipore 05-386 RRID: AB_11212640), and human EGFR (described in ref. ).

Techniques: Amplification, Labeling, Staining