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. 2016 Aug 10;291(39):20787–20797. doi: 10.1074/jbc.M116.723015

FIGURE 8.

FIGURE 8.

Nuclear and monomeric NPM1 protects against mHTT toxicity. A, N2a cells were either left untransfected (Un) or transfected with EGFP, Q138-GFP, or Q138-RFP. The cells were lysed 24, 48, or 72 h later and subjected to cross-linking as described under “Experimental Procedures.” The blots were probed for endogenous NPM1 and then reprobed with GFP and RFP antibodies. B, cortical neurons were transfected with either Q138-GFP (top panel) or Q138-RFP (bottom panel), and the cells were fixed 24 h later. Immunocytochemistry was performed by co-staining with NPM1 and either GFP or RFP antibodies. C, cortical neurons transfected in a 1:2 ratio with either EGFP and a control vector (pK3HA) or Q138-RFP and EGFP, NPM1, NESM, NLSD, or NPM1121–294. The cells were fixed 24 h later, and immunocytochemistry was performed with RFP and GFP antibodies. Viability was quantified by RFP or GFP fluorescence and DAPI staining. ***, p < 0.001 as compared with EGFP/control transfected cells; ###, p < 0.001 as compared with Q138-RFP/EGFP transfected cells (n = 3). Ctrl, control.