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. 2019 Jul 22;17:80. doi: 10.1186/s12964-019-0395-6

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© The Author(s). 2019

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Fig. 3 — Characterizing the expression of CD44-ICD in PCa cell lines. a. Immunoblotting analysis with an antibody to CD44-ICD (top panel) and GAPDH (bottom panel). Equal amounts of protein lysates (40 μg) made from PC3 (C) – cytoplasmic fraction (lane 1), PC3 (N) – nuclear fraction (lane 2), LNCaP (C) – cytoplasmic fraction (lane 3), LNCaP (N) – nuclear fraction (lane 4), PCa2b (C) – cytoplasmic fraction (lane 5), PCa2b (N) – nuclear fraction (lane 6) were immunoblotted with CD44-ICD antibody to detect cytoplasmic and nuclear levels. b. IB analysis of total (T), cytoplasmic (C) and nuclear (N) lysates (20μg) from PC3 cells with an antibody to CD44-ICD (~ 16.5 kDa). c. IB analysis of total cellular lysates (40μg) from PC3 (lane 1), LNCaP (lane 2) and PCa2b (lane 3) cells were immunoblotted for antibody to CD44-ICD. GAPDH was used as a loading control for immunoblotting analysis (A, lane 1, 3 & 5 and B, lane 1 & 2 and C). Nucleoporin (NP) was used as a loading control for nuclear (N) lysates (A, lane 2, 4 & 6, and B, lane 3). The results represent one of three separate experiments performed with the same results. Two asterisks (**) represents the 25 kDa CD44 extracellular truncation (CD44-EXT) while one asterisk (*) represents the 20 kDa CD44 extracellular truncation (CD44-EXT)