Absence of cell surface ROR1 in normal adult and pediatric tissues.

<p>(<b>A</b>) Immunohistochemical analysis of FFPE slides of a selection of normal adult tissues (top, heart; middle, cerebrum; bottom, adipocytes) probed with goat anti-human ROR1 pAbs revealed cytoplasmic staining in cardiomyocytes and neurons and plasma membrane staining in adipocytes. (Magnification 20×). (<b>B</b>) A protein lysate (5 µg/lane) from Burkitt lymphoma cell line CA-46 was compared to protein lysates (20 µg/lane) from 14 normal pediatric tissues by Western blotting using polyclonal goat anti-human ROR1 pAbs followed by HRP-conjugated donkey anti-goat IgG pAbs. None of the normal pediatric tissues revealed a firm ∼120 kDa band indicative of cell surface ROR1 expression although a faint ∼120 kDa band was detected in heart (Hrt), kidney (Kid), and uterus (Ute). Bands of higher and lower molecular weight are discussed in the text. The previously described <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0052655#pone.0052655-Yang1" target="_blank">[25]</a> recombinant fusion protein Fc-ROR1, which runs as ∼75-kDa band under reducing conditions, was used as additional control. The membrane was stripped and probed with rabbit anti-human GAPDH (∼36 kDa) followed by HRP-conjugated goat anti-rabbit IgG pAbs to confirm uniform loading of protein lysates. Abbreviations: Liv, liver; Lun, lung; Pan, pancreas; Ova, ovary; B, CD19+ B cells; Cbr, cerebrum; Cbe, cerebellum; Hrt, heart; Spl, spleen; Bla, bladder; Int, small intestine; Sto, stomach; Kid, kidney; Ute, uterus; Mus, skeletal muscle.