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Fig. 5. MATR3 is a substrate for CAPN1, and the pathogenic S85C mutation renders it resistant to degradation. Transfection of HEK293T cells with calpain expressing constructs resulted in robust expression of CAPN1 (A) and <t>CAPN2</t> (B) compared to controls. (C and D) CAPN1 but not CAPN2 overexpression resulted in MATR3 degradation (n = 3; *P = 0.010; one-way ANOVA with Tukey’s post hoc test). (E and F) While exogenous FLAG-MATR3(WT) is susceptible to cleavage by CAPN1 (n = 3; *P = 0.036; ns, not significant; one-way ANOVA with Tukey’s post hoc test), the pathogenic S85C mutation is resistant (n = 3; one- way ANOVA with Tukey’s post hoc test).
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Fig. 5. MATR3 is a substrate for CAPN1, and the pathogenic S85C mutation renders it resistant to degradation. Transfection of HEK293T cells with calpain expressing constructs resulted in robust expression of CAPN1 (A) and CAPN2 (B) compared to controls. (C and D) CAPN1 but not CAPN2 overexpression resulted in MATR3 degradation (n = 3; *P = 0.010; one-way ANOVA with Tukey’s post hoc test). (E and F) While exogenous FLAG-MATR3(WT) is susceptible to cleavage by CAPN1 (n = 3; *P = 0.036; ns, not significant; one-way ANOVA with Tukey’s post hoc test), the pathogenic S85C mutation is resistant (n = 3; one- way ANOVA with Tukey’s post hoc test).

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Neuronal activity regulates Matrin 3 abundance and function in a calcium-dependent manner through calpain-mediated cleavage and calmodulin binding.

doi: 10.1073/pnas.2206217120

Figure Lengend Snippet: Fig. 5. MATR3 is a substrate for CAPN1, and the pathogenic S85C mutation renders it resistant to degradation. Transfection of HEK293T cells with calpain expressing constructs resulted in robust expression of CAPN1 (A) and CAPN2 (B) compared to controls. (C and D) CAPN1 but not CAPN2 overexpression resulted in MATR3 degradation (n = 3; *P = 0.010; one-way ANOVA with Tukey’s post hoc test). (E and F) While exogenous FLAG-MATR3(WT) is susceptible to cleavage by CAPN1 (n = 3; *P = 0.036; ns, not significant; one-way ANOVA with Tukey’s post hoc test), the pathogenic S85C mutation is resistant (n = 3; one- way ANOVA with Tukey’s post hoc test).

Article Snippet: CAPN1 (#60941) and CAPN2 (#60942) overexpression constructs were purchased from Addgene.

Techniques: Mutagenesis, Transfection, Expressing, Construct, Over Expression