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Figure 5 | BMC Cancer

Figure 5

From: Tracking human multiple myeloma xenografts in NOD-Rag-1/IL-2 receptor gamma chain-null mice with the novel biomarker AKAP-4

Figure 5

RT-PCR/Western blot analyses of AKAP-4 mRNA/protein expressions and. RT-PCR analysis of mouse tissues was performed to evaluate AKAP-4 expression. AKAP-4 was detected in bone marrow (BM), blood (BL) and spleen (SP) of mice injected with MM cell lines (U266, H929) and primary tumor cells, but not in tumor-free mice. Positive controls were RNA isolated from exponentially in vitro growing cell lines, or from primary cells after ficoll purification. For the evaluation of healthy mice, RNA isolated from human testis was used as a positive control. Analysis of β-actin transcript served to confirm RNA integrity. Negative controls were PCR performed without template (No T), or with RNA not subjected to the reverse transcription step (No RT). Proteins extracted from MM cells and mouse tissues were analyzed for the presence of AKAP-4 protein by Western blot. AKAP-4 protein was detected in MM cell lines (U266 and H929), primary MM cells, and in the bone marrow, blood and spleen of tumor-challenged mice, but not in healthy controls (tumor-free). Total protein extracts from human testis were used as a positive control for the analysis of tumor-free mice. The pictures are representative of comparable results independently obtained from 5 mice in each group.

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