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Fig. 6 | Acta Neuropathologica Communications

Fig. 6

From: RNA-recognition motif in Matrin-3 mediates neurodegeneration through interaction with hnRNPM

Fig. 6

hnRNPM physically interacts with MATR3 in mammalian cells. a C2C12 cells ectopically expressing WT and mutant FLAG-MATR3 showed colocalization of FLAG-MATR3 (green) and endogenous hnRNPM (red) in the nucleus and cytoplasm. White arrows point to sequestration of endogenous hnRNPM with FLAG-MATR3 in cytoplasmic granules. b Quantification of number of cytoplasmic granules per cell showed that cell expressing F115C and S85C mutant MATR3 had increased formation of cytoplasmic granules (n = 6 per group; One-way ANOVA). c Quantification of percentage cytoplasmic granules that are positive for hnRNPM. Cells ectopically expressing mutant MATR3 showed increased sequestration of hnRNPM in cytoplasmic granules compared to WT (n = 6 per group; One-way ANOVA). d Immunoblot of co-immunoprecipitation of FLAG-MATR3 from HEK293T cells expressing FLAG-MATR3 and V5-hnRNPM. Immunoprecipitation with FLAG antibody showed V5-hnRNPM pulled down with FLAG-MATR3 WT, F115C and S85C. RRM2 deletion disrupts interaction between FLAG-MATR3 and V5-hnRNPM. e Immunoblot of lysates immunoprecipitated with FLAG antibody and treated with RNase A. Treatment with RNase A decreases interaction between FLAG-MATR3 WT and V5-hnRNPM (left). Elution of V5-hnRNPM in the unbound fraction after RNaseA treatment (right). Error bars indicate S.E.M. *p < 0.05; **p < 0.01

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