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. 2020 Dec 9;15(12):e0242677. doi: 10.1371/journal.pone.0242677

Fig 4. Overexpressed Ty1-TbMORN1 is predominantly cytosolic.

Fig 4

(A) Schematic of the two-step fractionation scheme. (B) Immunoblots of fractions taken from control and Ty1-TbMORN1 overexpressing cells, using anti-BiP, anti-TbMORN1 and anti-GFP antibodies. Note that the membrane was cut into three strips for the immunoblot. Equal fractions (5%) were loaded in each lane. The overexpressed Ty1-TbMORN1 was predominantly extracted by digitonin and partitioned with the cytosolic GFP into the SN1 fraction (arrows 1). Very little of the remainder was subsequently extracted with non-ionic detergent into the SN2 fraction (arrows 2), with most partitioning into the cytoskeleton-associated P2 fraction. Three independent experiments were carried out using cells from three clones pooled together; an exemplary blot is shown. (C,D) Quantification of the immunoblots of the two-step fractionation. Data were compiled from three independent experiments, each using cells pooled from three separate clones. Bars show mean values + SD.