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. 2014 May 2;9(5):e96326. doi: 10.1371/journal.pone.0096326

Figure 2. Increasing the levels of individual CP subunits alone has no effect on the endogenous levels of the other subunit, while co-expressing HA-cpa or HA-cpaΔABD and cpb enhance synergistically the levels of both subunits.

Figure 2

(A) model by which Cpa and Cpb cap F-actin barbed ends via the interaction of α and β tentacles with actin. (B) western blot on protein extracts from wing discs expressing UAS-mCD8-GFP (lane 1) or UAS-mCD8-GFP and UAS-HA-cpa89E (lane 2) or UAS-HA-cpa89E and UAS-cpb7 (lane 3) or UAS-mCD8-GFP and UAS-HA-cpaΔABD (lane 4) or UAS-HA-cpaΔABD and UAS-cpb7 (lane 5) under sd-Gal4 control, blotted with anti-HA (middle panel) and anti-H3 (lower panel). The means for lane 1 is 0, for lane 2 is 0.6250, for lane 3 is 2, for lane 4 is 0.0667, for lane 5 is 1.300. Error bars indicate s.e.m.. P<0.0092 for comparison of lanes 2 and 3 and of lanes 4 and 5. (B) western blot on protein extracts from wing discs expressing UAS-mCD8-GFP (lane 1) or UAS-mCD8-GFP and UAS-cpb7 (lane 2) or UAS-cpb7 and UAS-HA-cpa89E (lane 3) under sd-Gal4 control, blotted with anti-Cpb (middle panel) and anti-H3 (lower panel). The upper panels in B and C represent a quantification of relative (B) HA or (C) Cpb intensity signals for each genetic combination, measured by 4 independent blots. The means for lane 1 is 0.1088, for lane 2 is 0.5699, for lane 3 is 0.7982. Error bars indicate s.e.m. P<0.0182 for comparison of lanes 2 and 3. (D–D″ to I–I″) optical cross sections through distal epithelia of third instar wing imaginal discs with apical sides up and posterior sides to the left in which hh-Gal4 drives (D–D″) UAS-cpb7 and one copy of UAS-mCD8-GFP (green in D) or (E–E″) UAS-HA-cpa89E and two copies of UAS-mCD8-GFP (green in E) or (F–F″) UAS-HA-cpa89E, UAS-cpb7 and one copy of UAS-mCD8-GFP (green in F) or (G–G″) UAS-HA-cpaΔABD and two copies of UAS-mCD8-GFP (green in G) or (H–H″) UAS-HA-cpaΔABD, UAS-cpb7 and one copy of UAS-mCD8-GFP (green in H) or (I–I″) UAS-cpbL262R, UAS-HA-cpaΔABD and one copy of UAS-mCD8-GFP (green in I). Discs are stained with anti-Cpb (Cyan blue) and (D–D″) anti-Cpa (magenta) or (E–E″ to I–I″) anti-HA (magenta), which reveals (E–E″ and F–F″) HA-cpa89E or (G–G″ to I–I″) HA-cpaΔABD expression. The scale bars represent 15 µm.