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. 2022 Feb 17;13:931. doi: 10.1038/s41467-022-28613-0

Fig. 4. Kansl1+/− mice exhibit autophagic defects and impaired neuronal and cardiac functions.

Fig. 4

a, c Representative immunofluorescence images of hippocampus CA1 regions (a) and hearts (c) with LC3B antibody (red). Nuclei are labeled with DAPI (blue). Scale bar, 10 μm. b, d Quantification of LC3B puncta intensity in (a) and (c). LC3B puncta intensity in Kansl1+/+ was normalized to ‘1’ (n = 3 mice in each group). e Electron microscopy images of hippocampus CA1 regions and hearts. N, nucleus. Arrows indicate autophagosomes. Scale bar, 500 nm. f Violin plot showing the number of autophagosomes per μm2 in (e). n = 22, 14, 18, 21 cells from 3 independent experiments. g Representative immunofluorescence images of hippocampus CA1 regions and hearts with Stx17 antibody (green). Nuclei are labeled with DAPI (blue). Scale bar, 10 μm. h Quantification of Stx17 intensity in (g). Stx17 intensity in Kansl1+/+ was normalized to ‘1’ (n = 6, 8, 3, 3 mice from left to right). i Novel object recognition test. Discrimination index was calculated as the ratio of time spent exploring the novel object versus both the novel object and the familiar object in the choice trial. n = (10 Kansl1+/+ mice and 10 Kansl1+/− mice). j Morris water maze test. Mean escape latencies time to find the platform. n = 10 Kansl1+/+ mice and 10 Kansl1+/− mice. k Representative images of Nissl staining for whole brain of Kansl1+/+ and Kansl1+/− mice. Scale bar, 500 μm. l Representative images of dendritic spine of hippocampus CA1 regions visualized with Golgi-cox staining. Scale bar, 5 μm. m Quantification of spine density in (l). n = 29, 21 (from 3 Kansl1+/+ and 2 Kansl1+/− mice). n Representative images of echocardiograms on mice. Scale bar, 100 ms. o Echocardiographic analyses of (n). Left, ejection fraction. Right, fractional shortening. n = (24 Kansl1+/+ mice and 21 Kansl1+/− mice). Mice tested above were about 5 months old. Source data are provided as a Source data file. All data are means ± SEM. b, d, f, h, i, m, o Two-tailed Student’s t-tests; j by two-way ANOVA test with Dunnett’s multiple hoc test.