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. 2023 Jul 4;15:1155630. doi: 10.3389/fnagi.2023.1155630

FIGURE 6.

FIGURE 6

(A–D) Western blot analysis of total brain lysate of WT-C57BL/6J male mice subjected to intra-peritoneal TMG injection for 1 month. The blots were probed for panel (A) O-GlcNAc, OGT, OGA, α-Tubulin (B) p-ERK 1/2, Total ERK 1, and APP. (C) Densitometry plot of ERK 1/2 phosphorylation normalized to total ERK 1 and (D) APP normalized to α-Tubulin. The dots represent the number of mice (n = 4 for control and n = 5 for TMG injected mice). Statistical significance was measured using un-paired-t-test analysis and p-value is indicated on the plots. (E–K) Western blot analysis of total brain lysate of WT-C57BL/6J male mice subjected to intra-peritoneal TMG injection for 6 months. The blots were probed for panel (E) O-GlcNAc, OGT, OGA, α-Tubulin (F) p-ERK 1/2, Total ERK 1, and APP (I) p-MEK 1/2, MEK 1/2, Dusp4 and α-Tubulin. Densitometry plot of panel (G) ERK 1/2 phosphorylation normalized to total ERK 1 (H) APP normalized to α-Tubulin. (J) MEK 1/2 phosphorylation normalized to total MEK 1/2 and (K) DUSP4 normalized to α-tubulin. The dots represent the number of mice (n = 4 for control and n = 5 for TMG injected mice). Statistical significance was measured using un-paired-t-test analysis and p-value is indicated on the plots. (L,M) Western blot analysis of SH-SY5Y neuroblastoma cells subjected to long term OGA inhibition with TMG treatment. The blots were probed for panel (L) O-GlcNAc, OGT, OGA, APP, α-Tubulin. (M) Densitometry plot of APP normalized to α-Tubulin. The dots represent the number of the number of experimental trials (n). Statistical significance was measured using paired-t-test analysis and p-value is indicated on the plots. **Indicates p-values that are significant and less than 0.01 (p < 0.01).