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. 2018 Feb 8;9:210. doi: 10.3389/fimmu.2018.00210

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Copyright © 2018 Wong, Oikonomou, Paolicelli, De Luca, Pariano, Fric, Tay, Ricciardi-Castagnoli and Zelante.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Death-associated protein kinase 1 (DAPK1) expression during LAP is not controlled by Leucine-rich repeat kinase 2 (LRRK2). (A) Protein expression of DAPK1 in A. fumigatus swollen conidia-stimulated D1 cells. For densitometry analysis, band densities for DAPK1 were normalized to the band density of the housekeeping protein, β-actin, after which the density of the DAPK1 band of the treated sample was compared with that of the untreated sample. Data are representative of three independent experiments. (B) Immunofluorescent stanining of DAPK1 and brightfield microscopy of Aspergillus fumigatus swollen conidia-stimulated D1 cells. Signal intensity of the DAPK1 was quantified and displayed as mean fluorescence signal ± SD of three biological replicates. Brightfield imaging of the same cell is displayed with arrows indicating the presence of A. fumigatus swollen conidia in the cell. (C) Knockdown of LRRK2 in D1 cells was assessed by western blot. Data are represented as mean ± SD of three biological replicates and statistical significance determined by two way ANOVA (D) mRNA expression of DAPK1 in fungal-treated D1 cells that have been silenced for LRRK2. Data are displayed as the mean mRNA expression ± SD of three biological replicates and statistical significance determined by two way ANOVA. Differences found to be statistically significant are indicated (*, p < 0.05; **, p < 0.01). Abbreviations: A. fumigatus swollen conidia (A-sw); Untreated (Unt); D1 cells transduced with non-targeting control shRNA (Ctrl).