Fig. 4.

The SEC-MAP detection was verified by Western blot (A), FACS (B) and quantitative real-time PCR (C). The line plots from SEC-MAP and the histograms from FACS show the expression of CD markers (CD22 and CD44) in the cell lines that were positive (solid line) and negative (dashed) for their respective proteins. The line plots show 24 SEC fractions (x axis) and the MFI of PE-labeled streptavidin (y axis). The protein entities of CD22 and CD44 are marked by vertical lines. The histograms from FACS show the MFI of fluorescent-conjugated antibodies against CD22 and CD44 (x axis) (A). The line plots from the SEC-MAP and Western blots (WB) show the expression of C-terminal-binding protein 2 (CTBP2), and SH2 domain-containing protein 1A (SH2D1A) in the cell lines that were positive (solid line, left WB bands) and negative (dashed, right WB bands) for their respective proteins. The line plot shows 24 SEC fractions (x axis) and the MFI of PE-labeled streptavidin (y axis). The protein entities of CTBP2 and SH2D1A are marked by vertical lines (B). Quantitative RT-PCR was used to test the mRNA expression. The ΔCq method was used for data normalization. Hypoxanthine-guanine phosphoribosyltransferase (HPRT1) and beta-glucuronidase (GUSB) were used as housekeeping genes. The line plots from the SEC-MAP show the STAT5A expression in the cell lines that were positive (solid line) and negative (dashed) for their respective proteins. The line plots show 24 SEC fractions (x axis) and the MFI of PE-labeled streptavidin (y axis). The protein entities of STAT5A are marked by vertical lines (C).