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. 2019 Nov 22;20(23):5866. doi: 10.3390/ijms20235866

Figure 2.

Figure 2

Pgrn protein content and secretion in murine TDP-43 gain-of-function (GOF) cells. (a) Representative immunofluorescence images showing the subcellular localization of the recombinant GFP-TDP-43 full length and C-terminal fragments (GFP-TDP-35; GFP-TDP-25) in NSC-34 cells. DAPI (blue) was used for nuclei staining. Scale bar: 10 µm. (b) Representative WB images of Poldip3 protein isoforms (α and β) in NSC-34 cells knocked-down for Tdp-43 (siTdp-43) or expressing GFP-TDP-43, GFP-TDP-35 and GFP-TDP-25 constructs as indicated. Immunoblots with anti-GFP and anti-Tdp-43 antibodies were performed to assess TDP-43 over-expression and gene silencing efficiencies, respectively. Upon GFP-TDP-43 over-expression, autoregulation of the endogenous Tdp-43 protein could be observed. αTubulin was used for sample normalization. (c) Representative WB images and densitometric analyses of Pgrn protein content in cell lysates (CELL) and conditioned media (CM) of NSC-34 cells transfected as indicated. αTubulin was used for sample normalization and as non-secreted control protein in CM (mean ± s.e.m.; n = 3 independent experiments; one-way ANOVA and Bonferroni’s multiple comparison post hoc test) *, residual signal from previous anti-GFP hybridization.