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. 2013 Oct 21;14(10):21114–21139. doi: 10.3390/ijms141021114

Figure 9.

Figure 9

Quantitative-PCR analysis of NGF expression in F11 cells after 72 h DADLE treatment following knockdown of the Oprd1 gene. Following transfection with Oprd1 siRNA (cocktail of three siRNAs, 500 nM each) and scrambled siRNA (1.5 μM), 0.5 million cells/60-mm dishes were differentiated with 0.5 mM db-cAMP and 50 ng/mL NGF for 72 h in medium supplemented with or without 1 μM DADLE. After 72 h, cells were harvested for total RNA isolation. Reverse transcription was performed with 2 μg of total RNA for each sample. PCR was performed for rat NGF, mouse Oprd1 and GAPDH. (A) Representative semi-quantitative PCR products that were run on a 3% agarose gel and stained with ethidium bromide; (B) Relative fold change of mouse Oprd1 as compared to GAPDH obtained from quantitative PCR. The data is normalized to one for the no siRNA treatment. Quantitative PCR was run using cDNA samples pooled from two independent runs. Data is from one PCR run done in replicates of four for each treatment; (C) Table showing the percent decrease of mRNA levels of NGF and Oprd1 following Oprd1 knockdown.