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Fig. 4. Sec 31B is widely expressed in tissues and in cultured cells. (A) Northern blots identify the predominant Sec31B mRNA as a series of two or three closely spaced bands centered at 6.5 Kb in all tissues examined, but heavily expressed in testis. Faint bands can also be noticed at approximately 1.0 kb and over 11kb, the later presumably representing traces of genomic contamination. Analysis by dot-blotting of a multiple tissue array confirms abundant expression in testis as well as in cerebellum, thymus, lymph node, pituitary gland and, to a lesser extent, in all tissues on the array. A complete key to the dot-blot array is presented in supplementary material Table S1. (B) Western blots of a series of cultured cell lines using the rabbit IgG 2013 antibody, compared with the same blots probed with the antibody against Sec31A. Preimmune sera shows only a prominent non-specific band at approximately 80 kDa. This non-specific band is barely detectable with the affinity-purified IgG 2013, it demonstrates reactivity with Sec31B-F at 129 kDa as well as with a number of smaller bands. Notice that there is no correspondence of the bands reacting with IgG 2013 and the Sec31A antibody. (C) Cell- and tissue-extracts prepared from rat brain and testis were western blotted with anti-Sec31B antibodies Mab 1G10 and IgY 1871. The control lanes (ctrl) depict the reactivity present in non-immune hybridoma medium against MDCK cells (for Mab 1G10) or in the pre-immune IgY (for IgY 1871). Notice the presence of a 129 kDa band with both antibodies (Sec31B-F) and that only Mab 1G10 reacts with a prominent 53 kDa band (Sec31B-T). The star marks the position of the faint non-specific band just below the band for Sec31B-T.
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