Fig 1: MICAL1 reactivation in a short window rapidly disrupts cued fear memory. (A) Schematic of the experimental design. (B-E) The effect of MICAL1 reactivation by recombinant mouse SEMA3A on fear memory retrieval was examined. Injection of recombinant mouse SEMA3A into the BLA at (B) 0.5 h, but not at (C) 1 h, (D) 2 h and (E) 24 h post-conditioning, significantly attenuated freezing behavior on retrieval (n = 8 mice per group). Two-way ANOVA with Bonferroni's multiple comparisons post hoc tests (left) or two-tailed, unpaired t tests (right) (B-E). ∗p < 0.05. Data are presented as the mean ± SEM.
Fig 2: Changes in Sema3A protein content in each group. A: WB showing Sema3A expression in RGCs cultured for 3 d, 7 d, or 10 d. B: The expression of Sema3A in cultured RGCs transfected with miR-30b mimic, inhibitor, miRNA NC, or Sema3A siRNA at 120-140 h (n=3). *p<0.05 compared to control siRNA.
Fig 3: The effects of exogenous Sema3A on axon and dendrite differentiation in RGCs. A: Fc-Sema3A (0.5 μg/ml) treatment for 72 h promoted the formation of RGC dendrites and inhibited axon growth, as demonstrated by immunostaining for neurites and dendrites with III-tubulin and the dendrite marker MAP2 antibodies, respectively. Filled and empty arrows denote axons and dendrites, respectively. The scale bar is 40 μm. B: The lengths of the longest axon and dendrite per RGC were measured after culture in Fc-Sema3A (0.5 μg/ml) after 72 h (mean±SD; 40-53 cells per treatment). C, D: The numbers of RGCs with different numbers of axons and dendrites were measured following treatment with Fc-Sema3A (1 μg/ml or 0.5 μg/ml) after 72 h. E: The percentage of bipolar cells was measured following treatment with Fc-Sema3A (1 μg/ml) after 72 h (41-53 cells per treatment). *p<0.05, **p<0.01.
Fig 4: Changes in RGC axons and dendrites after transfection with Sema3A siRNA. A: Treatment with siRNA Sema3A and control siRNA for 120 h promoted the growth of RGC axons and inhibited the growth of dendrites. Scale bar, 40 μm. B: The longest axon and dendrite per RGC were measured after transfection with Sema3A siRNA at 120 h of culture. C: The percentage of bipolar cells was measured in Sema3A siRNA-treated RGCs after 120 h (mean ± SD; 37-71 cells per treatment, *p<0.05 compared to control siRNA).
Fig 5: Differences in p-PKA, GSK-3β (S9), and CRMP2 (T514) expression among the control, miR-30b mimic, Fc-Sema3A, and miRNA NC groups. A: WB showing p-PKA, GSK-3β (S9), and CRMP (T514) expression after transfection with miR-30b mimic or 1μg/mL Fc-Sema3A. B: Fluorescence intensity of CRMP2 (T514) in the different groups after 60 h of culture (n=3 cultures; p<0.05, **P<0.01 compared to miRNA NC or control; #p<0.05, ##p<0.01 compared to the alternative dose).
Supplier Page from R&D Systems, a Bio-Techne Brand for Semaphorin 3A Fc Chimera Protein
Available conjugates: Sizes Available: 25 ug