Degradation of TRPML1 in Neurons Reduces Neuron Survival in Transient Global Cerebral Ischemia

Figure 1

Proteasome-mediated degradation of TRPML1 of hippocampal neurons in ischemia. (a–d) Representative immunoblots of TRPML1 from cultured hippocampal neurons (a, ) or astrocytes (c, ) 0.5 or 1 hrs after OGD. (b) or (d) was the statistics for (a) or (c). (e, f) Immunoblots of analysis of the indicated marker expression in sorted neurons (e) and astrocytes (f) (GFAP: astrocyte marker, NeuN: neuron marker). (g–j) Representative immunoblots of TRPML1 from sorted neurons (g, ) or astrocytes (i, ) 40 min after tBCCAo. (h) or (j) was the statistics for (g) or (i). (k) Quantitative PCR analysis of TRPML1 mRNA levels in hippocampal neurons 1 hr after OGD (). (l, m) Representative immunoblots of TRPML1 from cultured hippocampal neurons () incubated with/without calpeptin (10 μM), DEVD (10 μM), or MG132 (5 μM). (m) was the statistics for (l). β-Actin serves as a loading control. All data were presented as mean ± SEM. Comparisons between groups for statistical significance were performed with one-way analysis of variance (ANOVA) with Tukey’s post hoc test (b, d, and k), Student’s t-test, two tailed (h, j), or two-way ANOVA with Bonferroni post hoc test (m). , versus Ctrl.

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