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. 2014 Feb 12;34(7):2413–2421. doi: 10.1523/JNEUROSCI.3226-13.2014

Figure 2.

Figure 2.

High concentrations of Coronin 6 at the NMJ. A, B, Specificity of Coronin 6 polyclonal antibody. Rabbit Coronin 6 polyclonal antibody recognized both the overexpressed Coronin 6 (Coro6) and endogenous Coronin 6 in adult rat muscle (Muscle) (A, left). This recognition was blocked by preincubation with the peptide antigen (A, right). Western blot analysis of COS-7 cells with ectopic expression of His-tagged Coro1C and Coro6 using the Coronin 6 polyclonal antibody (B). C, Coronin 6 protein expression in rat muscle during development was determined by Western blotting. D, Western blot analyses of Coronin 6 (20 μg per lane) were performed using rat gastrocnemius muscle after sciatic nerve crushing. Actin served as a loading control. E, C2C12 myoblasts in growth media (GM) or myotubes in differentiation media for the indicated times (h) were lysed and subjected to Western blotting to examine Coronin 6 protein expression. F, Coronin 6 was concentrated at the motor endplates during development (postnatal stages, from P1 to P21). Rat skeletal muscle sections were costained with antibody against Coronin 6 and AlexaFluor-555-conjugated α-BTX to visualize AChRs. Scale bar, 10 μm. Arrowheads indicate Coronin 6 staining colocalized with AChRs; arrows indicate Coronin 6 not colocalized with AChRs. G, Coronin 6 was localized at the postsynaptic muscle membrane of gastrocnemius muscle after nerve denervation. Scale bar, 10 μm. H, The immunoreactivity of Coronin 6 antibody was abolished by preabsorption with its peptide antigen. Scale bar, 10 μm.