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Fig. 4 | Acta Neuropathologica Communications

Fig. 4

From: Methylene blue inhibits Caspase-6 activity, and reverses Caspase-6-induced cognitive impairment and neuroinflammation in aged mice

Fig. 4

MB inhibits Casp6 activity in the brains of aged mice. a Representative two photon images of Casp6-FLICA stained CA1 neurons in acute brain slices from vehicle-treated KI/Cre (ACL/G n = 1, ACL n = 2), MB-treated KI/Cre (ACL n = 3), or WT/WT (n = 3) mice, and z-VEID-FMK-treated KI/Cre (ACL/G n = 1, ACL n = 2) mice. IR-DIC: infrared differential interference contrast, TFM: two-photon microscope. b Quantification of the numbers of FLICA positive neurons per mm along the pyramidal neuron. Data represents mean ± s.e.m. Statistical evaluations were done with one-way ANOVA (p = 0.0026) followed by Dunnett’s post-hoc analysis. ****p < 0.01 vs vehicle-treated WT/WT. c Western blotting and d quantification of Casp6 in hippocampus or cortex from vehicle- (n = 6) or MB-treated (n = 5) ACL/G KI/Cre mice. Uncleavable Casp6D23,179,193A recombinant protein, recombinant WT Casp6 protein, which has lost its pro-domain during purification, and Casp6 knock out mouse proteins were used as controls. Data shown as mean and s.e.m. Each symbol represents data from one mouse. e&f Representative micrographs of immunopositive Tub∆Casp6 in the SO, PCL, SR, and SLM CA1 regions (e), and in corpus callosum, fimbria, and fornix (f) of vehicle-treated ACL/G KI/Cre (n = 5) or WT/WT (n = 3) mice, and MB-treated ACL/G KI/Cre (n = 2) mice. Bar = 50 μm. dhc: hippocampal commissure, df: dorsal fornix; arrows indicate immunopositive neurites

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