Detection of JUP isoforms in macrophage-like cells.

To differentiate THP1 into macrophage-like cells, cells were stimulated by PMA for 1, 2, 3 and 4 days. JUP isoforms were analyzed by Western blotting using anti-JUP antibodies LS-C77817 (a), ab134558 (b), CP2971 (c) and CM1111 antibody (d). 200, 100, 50 and 25 ng of recombinant GST-tagged JUP was used as positive control (lanes 1–4, respectively). Lane 5 contains lysates of undifferentiated THP1 cells (monocytes) and lanes 6–9 contain lysates of differentiated THP1 cells (macrophages) after 1, 2, 3 and 4 days of differentiation (note opposite order of loading in panel b)). β-actin was employed as a loading control. CP2971 reacts only weakly with recombinant JUP on Western blots (no visible reaction detected here).