Figure 2

Signal amplification in IF with FT-GO. (a–l) IF for Rbfox3 using a direct (a–d), indirect (e–h) and FT-GO method (i–l) in layer 2/3 neurons of the mouse S1 (n = 3 animals for each condition). Brain sections were reacted with an anti-Rbfox3 Ab at dilutions of 1:100 (a,e,i), 1:1000 (b,f,j), 1;10,000 (c,g,k) and 1:100,000 (d,h,l). CF488A tyramide is used for color development in the FT-GO method. Images are acquired with the same parameters for comparisons. (m) Histograms representing fluorescence signal intensities (AU) of Rbfox3-positive cells in the layer 2/3 of S1 (n = 630 cells, primary [pAb] 1:100, direct method; n = 644 cells, pAb 1:100, indirect method; n = 686 cells, pAb 1:100, FT-GO; n = 622 cells, pAb 1:1000, direct method; n = 698 cells, pAb 1:1000, indirect method; n = 758 cells, pAb 1:1000, FT-GO; n = 620 cells, pAb 1:10,000, direct method; n = 639 cells, pAb 1:10,000, indirect method; n = 728 cells, pAb 1:10,000, FT-GO; n = 650 cells, pAb 1:100,000, direct method; n = 619 cells, pAb 1:100,000, indirect method; n = 686 cells, pAb 1:100,000, FT-GO; n = 3 animals for each condition, pAb 1:100; H = 1737, df = 2, P < 0.0001, Kruskal–Wallis test, pAb 1:1000, H = 1832, df = 2, P < 0.0001, Kruskal–Wallis test, pAb 1:10,000; H = 1744, df = 2, P < 0.0001, Kruskal–Wallis test, pAb 1:100,000; H = 1726, df = 2, P < 0.0001, Kruskal–Wallis test, ***P < 0.0001; Dunn’s post hoc test). Data are represented as means ± SDs. Scale bar: 25 µm.