Fig. 2
The localization of calpain 5 in the olfactory epithelium. a, b, CAPN5 mRNA in situ hybridization images (left panels, bright field) in the presence of an antisense (a) or sense (b) probe and GFP images (right panels, green; a, b) in the same areas. Note that the strong signals were detected along the duct from Bowman’s gland (indicated by +). k and s indicate the knob and soma of ORNs, respectively. c, Brightness values that declined in the presence of dye in (a, b) are defined as signals are and shown as 8-bit brightness values in an arbitrary unit (a.u.; 28 as the max). Statistics; * P < 0.001; the T-values (df) are 11.7 (38), 4.61 (38), and 11.0 (38) for SC, s, and LP (blue versus white), respectively. ROIs (20 total). Signals were compared between soma and the other olfactory regions; # P < 0.001; the T-values (df) are 7.73 (38) and 9.21 (38) for SC and LP, respectively. d-g, Immunohistochemistry images (red) with nuclear staining with 4′,6-diamidino-2-phenylindole (DAPI, blue) and GFP fluorescence (green) at low magnification (d, e) and at high magnification (f, g) in the olfactory regions in the presence (d, f) or absence (e, g) of the anti-CAPN5 primary antibody (1st Ab). h-k, Verification of CAPN5 antibodies in the other brain regions in the presence (h, j) or absence (i, k) of 1st Ab. CAPN5-IR (red), DAPI (blue) and GFP fluorescence (green). fim, fimbria. The GFP signals represent the OMP-expressing ORNs and projections. Only the merged images are presented in (h) to (k). l, m, The CAPN5-IR signal intensities (a.u.; 28 as max) for olfactory tissues (l) and the central nervous system (m). Statistics; * P < 0.001; the T-values (df) are 31.9 (100), 60.2 (114), 18.4 (115), 34.9 (99), 97.1 (99), 18.3 (98), 32.4 (105) and 37.8 (114) for SC, s, LP, Ctx, Hip, LGB, CPu and Pir (red versus black), respectively. To compare CAPN5-IRs between soma and other tissue regions, the same ROIs were statistically analyzed; # P < 0.001; the T-values (df) are 31.1 (112), 10.8 (124), 14.2 (112), 26.9 (111), 9.8 (111), 17.8 (115) and 17.8 (124) for s in (f) against SC, LP, Ctx, Hip, LGB, CPu and Pir (red versus red), respectively. ROIs (50–63 total) of the experiments with or without the 1st antibody. Representative ROIs are shown in open rectangles: 30 μm2 in (a, f); otherwise, 1400 μm2 in (h) and (j). Scales: 30 μm in (a, b, f, g); otherwise, 200 μm. Dotted lines in (a, b, f, g) indicate the basal membrane separating OE and LP