movie S3.mp4

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movie S2.mp4

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movie S6.mp4

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movie S5.mp4

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movie S4.mp4

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movie S1.mp4

Visualization

Protective effects of GlcNAc-Sal on OGD-R-induced cytotoxicity in HT22 cells.

<p>Cell viability was determined by the MTT assay and LDH release assay. (A) HT22 cells were exposed to OGD insults of different durations followed by 24 h of reperfusion. (B) HT22 cells were exposed to different concentrations of GlcNAc-Sal for 24 h. The effect of GlcNAc-Sal on the viability of HT22 cells exposed to OGD-R stimulation was analyzed by the MTT assay (C) and LDH release assay (D). (E) Morphological alterations in HT22 cells. All data were expressed as mean ±SEM of three independent experiments performed in triplicate. ^##<i>p</i><0.01, ^###<i>p</i><0.001 vs. control; *<i>p</i><0.05, **<i>p</i><0.01 vs. OGD-R alone.</p

GlcNAc-Sal inhibits NO production, NO-induced damage and the expression of iNOS in OGD-R-treated HT22 cells.

<p>(A) The Griess reagent was used to evaluate the content of NO in the culture supernatant after OGD-R treatment in the presence and absence of GlcNAc-Sal. Quantitative assessment was performed using NaNO₂ as a standard. The effect of GlcNAc-Sal on the cell viability of HT22 cells exposed to SNP stimulation was analyzed by the MTT assay (B) and LDH release assay (C). (D) The morphological features of apoptosis in HT22 cells were detected by Hoechst 33342 staining. Arrowheads indicted apoptosis cells. (E) The rates of nuclear condensation were calculated in comparison to treatment with SNP alone. (F) Effect of GlcNAc-Sal on the expression of the iNOS protein induced by OGD-R treatment. GAPDH was used for normalization. (G) The intensity of bands was quantified by densitometric analysis. The data are expressed as mean ± SEM from three independent experiments. ##<i>p</i><0.01, ###<i>p</i><0.001 vs. control group; *<i>p</i><0.05, **<i>p</i><0.01, ***<i>p</i><0.001 vs. OGD-R group or SNP group.</p

Effects of GlcNAc-Sal on the protein levels of Bcl-2, Bax, caspase-3 and PARP in OGD-R-treated HT22 cells.

<p>(A) The expression levels of Bcl-2, Bax, caspase-3 and PARP were analyzed by Western blotting. Anti-GAPDH antibody was used for normalization. (B) The intensity of bands was quantified by densitometric analysis. All values represent mean ±SEM of three independent experiments. ^#<i>p</i><0.05, ^##<i>p</i><0.01 vs. control; *<i>p</i><0.05, **<i>p</i><0.01 vs. OGD-R alone.</p

Chemical structure of salidroside and GlcNAc-Sal.

<p>(A) p-hydroxyphenethyl-β-D- glucoside. (B) 2-(4-Methoxyphenyl)ethyl-2-acetamido-2-deoxy-β-D-pyranoside.</p