Figure 3 | Scientific Reports

Figure 3

From: Photoactivated adenylyl cyclase (PAC) reveals novel mechanisms underlying cAMP-dependent axonal morphogenesis

Figure 3

Subcellular expression of PKA and Epac in the axonal region.

(ae) Left, Representative images of cultured granule cells at DIV 4, immunolabeled by antibodies against PKA catalytic α/β subunits (a, PKA cat), PKA regulatory domain I (b, PKA RI), PKA regulatory domain II α (c, PKA RII), Epac1 (d), or Epac2 (e). Axon was immunolabeled by anti-Tau antibody and a whole cell morphology (actin) was labelled by rhodamine phalloidin. Right, Magnified images of the boxed region in the left panels. Scale bar = 20 μm. (f) Quantifications of the subcellular distribution of PKA cat, PKA RI, PKA RII, Epac1 and Epac2 in branches. The ratio of fluorescence intensity in the axonal branches to that of the primary axonal shafts reveals that PKA localized more in primary axons than in axonal branches, whereas both Epac1 and Epac2 exhibited higher localization in branches. **p < 0.01 between indicated groups; Tukey’s test after one-way ANOVA, n = 10 cells for each group. (f) Quantifications of the subcellular distribution of PKA cat, PKA RI, PKA RII, Epac1 and Epac2 in growth cones. The ratio of the growth cone fluorescence intensity to that of the primary axonal shafts demonstrates the strong accumulation of Epac2 in growth cones. **p < 0.01 between indicated groups; Tukey’s test after one-way ANOVA, n = 10 cells for each group.

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