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B
D
Figure 4. Two levels of analysis of each gene RNA and protein. mRNA expression of each gene was assessed by qRT-PCR and normalized relative to GAPDH and expressed as 2-ΔCt (A). Abundance of each protein was assessed by CNIA and normalized relative to GAPDH abundance in each case (B). The Y axis of graphs (A) and (B) are expressed on a log scale. The variability of each mRNA and protein measurement in the analyzed population of patients with MM, measured as percentage coeffcient of variation (CV%). The threshold of statistical significance (*P<0.05) was determined as described in the Methods section (C). Spearman correlation coef- ficient for each mRNA/protein pair ranked by increasing P value (D).
Nano-scale protein quantification in multiple myeloma
Discussion
MM has been comprehensively studied at the DNA and RNA levels using high-throughput technologies such as microarrays for detecting copy number abnormalities, and gene expression profiling, and, more recently, next-gener- ation sequencing for DNA mutation analysis. MM sam-
ples after CD138+ separation are usually stored in buffers such as TRIZOL or RLT Plus, which preserve nucleic acids for subsequent use in genomic studies. While it is techni- cally possible to extract proteins from these buffers,23–25 the quantity of protein would not be sufficient for multiple WB to be carried out. In the classic WB, the amount of the purified plasma cells, even if all of it were available for the
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