Fig 1: In situ hybridization of lncRNA and mRNA in neuron cells. (A) Biotin-labeled lncRNA and digoxigenin-labeled mRNA probes are shown in green and red, respectively. LncRNA and mRNA are co-expressed in CA1 of the hippocampus. (B, C) The colocation effect of LncRNA (B) or mRNA (C) with MAP-2 labeled neuron cells indicated these correlations mainly happened in neuron cells of the hippocampus. LncRNA and mRNA were labeled by red fluorescent probes, and he neuron cells were marked using anti-MAP2 antibody and Alexa 488 conjugated anti-rabbit IgG. Scale bar: 50 µm.
Fig 2: CXCL5 localization in the hippocampus of rats. The brain tissues were then fixed in 4% paraformaldehyde in phosphate-buffered saline (PBS) for 24 h at 4 °C. Immunofluorescence analysis was applied to show CXCL5 localization in rat hippocampus. a, representative fluorescence images show the co-localization of CXCL5-positive staining and GFAP-positive staining. b, representative fluorescence images show the co-localization of CXCL5-positive staining and MAP2-positive staining. Arrows indicate the cells with double staining
Fig 3: 2JY-OBZ4 ameliorated human oligomeric Aß42 induced synaptic loss in mice primary neuron. (A) CCK-8 assay of mice primary neuron incubated with various concentrations of 2JY-OBZ4 (0 nM, 125 nM, 250 nM, 500 nM, 1µM, 2 µM, 4 µM, 8 µM, 16 µM, 32 µM, 64 µM, 128 µM) for 24 h. n = 3 per group. p value significance is calculated from a one-way ANOVA, data are represented as mean ± SEM. *p < 0.05, compared to controls. (B) Western blots and (C, D) quantitative analysis for synaptophysin and PSD95 in mice primary neuron. MW Molecular weight. n = 3 per group. #p < 0.05 compared to controls, *p < 0.01, **p < 0.01, ****p < 0.0001 compared to the group pretreated with Aß and treated with DMSO (E–J) Immunofluorescence staining was used to measure the expression of synaptophysin and Map2 in primary neuron (scale bar: 50 µm). (K, L) Quantitative analysis of fluorescence intensity. n = 4 per group. p value significance is calculated from a one-way ANOVA, data are represented as mean ± SEM. #p < 0.05, #### p < 0.0001 compared to controls, *p < 0.01, ***p< 0.001 compared to the group pretreated with Aß and treated with DMSO.
Fig 4: Gata3 inhibition induces cell differentiation in 661W cells. (A) Strong Nestin (red) and weak Map2 (green) expression were observed in the 661W cells. In Gata3-silenced 661W cells, Nestin expression was decreased, and Map2 expression was observed. The white arrow indicates neurite outgrowth stained by Nestin, the white arrowhead indicates neurite outgrowth stained by Map2. (B) Gata3 inhibition decreased the viability of 661W cells. (C), Both Gata3 silencing and Brca1 silencing induce synaptophysin (red, cell cytoplasm) expression in 661W cells. Scale bars represent 20 µm. The asterisks indicate statistically significant differences (** p < 0.01).
Fig 5: Brca1 silencing induces differentiation of 661W cells. (A) Strong Nestin expression (red) and faint Map2 expression were observed in the cell cytoplasm of 661W cells. Brca1 was expressed at high levels in the cellular nuclei of 661W cells (green). The nuclei were stained with DAPI (blue). (B) Nestin expression was decreased in Brca1-silenced 661W cells, as evidenced by Western blot. (C) The relative expression levels of Nestin and Brca1 in 661W cells are presented as a histogram. (D) Brca1 silencing significantly increased the expression of Map2 (C/D) in 661W cells. (E) The relative protein expression levels of Map2 (A/B) and Map2 (C/D) are presented as histograms. Scale bars represent 10 µm. The asterisks indicate statistically significant differences (* p < 0.05, ** p < 0.01).
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