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Fig. 3 | Molecular Brain

Fig. 3

From: TMEM106B, a frontotemporal lobar dementia (FTLD) modifier, associates with FTD-3-linked CHMP2B, a complex of ESCRT-III

Fig. 3

T185 was sequestered more in CHMP2BIntron5-positive aggregates. a CHMP2BIntron5-cMyc (CHMP2BIn5-cMyc) was cotransfected with either Flag-tagged T185 or S185 into the HEK293T cells. Twenty-four to 48 h after transfection, the cell lysates were used for coimmunoprecipitation with an anti-flag antibody. Western blot analyses were performed with anti-cMyc or anti-flag antibodies. b The bar graph represents the percentages of interaction of T185 or S185 with CHMP2BIn5. For quantification, the band intensity of CHMP2BIn5 from the immunoprecipitation samples was normalized to that of either flag-tagged T185 or flag-S185. The values are presented as the mean ± SEM of three independent replicates. Student’s t-test, *p < 0.05. c Each flag-tagged TMEM106B SNP (T185 or S185) was cotransfected with CHMP2BIn5-cMyc into cultured cortical neurons. Twenty-four hours after transfection, the neurons were fixed and stained with an anti-flag antibody and anti-cMyc antibody. The arrows indicate the colocalization of CHMP2B with either T185 or S185. Scale bar: 20 μm. d The bar graph indicates the colocalization of CHMP2BIn5 with either the T185 or S185 isoform of TMEM106B. The coefficients of colocalization were measured and analyzed with the ImageJ program (Colocalization Indices). The values are presented as the mean ± SEM of three independent replicates. Student’s t-test; ***p < 0.001

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