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. 2020 Sep 4;133(17):jcs248880. doi: 10.1242/jcs.248880

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© 2020. Published by The Company of Biologists Ltd

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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Fig. 6. — Heightened Rac activation in MFN2-deficient dHL-60 cells is corrected by inducing a mitochondria–ER tether. (A) Western blot and (B) quantification determining the amount of phospho-PAK (pPAK) in dHL-60 cells treated with fMLP at indicated time points. L, protein ladder. (C) Western blot determining the amount of Rac-GTP and total Rac protein in dHL-60 cells treated with fMLP at indicated time points. (D) Quantification of Rac activation 5 min after stimulation with fMLP. (E) Immunofluorescence of F-actin and Rac-GTP in indicated cell lines 3 min after stimulation with fMLP. Arrows, direction of cell polarization. (F) Colocalization of Rac-GTP and F-actin. n>20. (G,H) Western blot (G) and quantification (H) determining the amount of pPAK in dHL-60 cells treated with fMLP at indicated time points. One representative result of three biological repeats is shown in A,C, and G. Data are pooled from three independent experiments in B, D and H. Error bars represent s.d. NS, non-significant; *P<0.05; **P<0.01 [unpaired t-test (B,D), and two-way ANOVA (H)]. Scale bar: 10 µm.