Part I (3 points): The researchers first need to establish the system that they

Question

Part I (3 points): The researchers first need to establish the system that they will use in their paper. The cell type they will use for their experiments is human umbilical vein endothelial cells (HUVEC). Based on the data in Figure 1A, what can you conclude about the inducibility of the iNOS gene and the VCAM-1 gene in HUVEC cells in response to cytokine stimulation? Explain your reasoning. Part Il (3 points): What can you conclude about the data in Figure 18? How do the data in panel B support your conclusion in Part I? Real-Time RT-PCR Pol lI ChIP s 16 1 INOS VCAM-1 iNOS VCAM-1 12 FIGURE 1. Effects of cytokines on iNOS and VCAM-I steady-state mRNA levels and transcription HUVEC. Primary HUVEC were stimulated with vehicle (negative control), TNF- (10 ng/ml), or a cytokine mixture (Cyt) of IFN-y(200 U/ml), IL-1 (5 ng/ml), and TNF- (10 ng/ml) for 4 h, after which RNA and chromatin extracts were isolated. A. Quantitative RT-PCR of human iNOS and VCAM-1 mRNA. Inducible NOS and VCAM-1 mRNA expressions were normalized to GAPDH mRNA level (in arbitrary units). Data represent the mean ± SEM of four independent experiments. B, ChIP assay using an antibody directed against RNA polymerase II (Pol II). IP DNA was analyzed by quantitative PCR using i OS and VCAMI promoter-specific primers. Data represent the mean ± D of triplicate measurements from one of four independent experiments with similar results. .p

Explanation / Answer

Based on the graph A, we can say that the VCAM-1 gene is induces successfully in the HUVEC calls while iNOS gene is not inducible in the cells as theres no response to cytokine stimulation.

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