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. 2014 Aug 14;10(8):e1004304. doi: 10.1371/journal.ppat.1004304

Figure 4. EBNA3C co-localizes with Pim-1 and facilitates its nuclear translocation.

Figure 4

A) 0.3 million HEK-293 cells plated on coverslips and transiently transfected with control vector, GFP-EBNA3C and Myc-Pim-1 expression vectors by using Lipofectamine 2000 transfection reagent. B) 10 million HEK-293 cells were transfected with Myc-Pim-1 and Flag-EBNA3C and subjected to sub-cellular fractionation assay. C) Bar diagram represents quantitation of co-localization in panel A. D) BJAB, BJAB10, LCL1 cells were plated on slides and air-dried. Ectopic and endogenous expressions of Pim-1 was detected using anti-Myc (9E10)-antibody (1∶200 dilution) and Pim-1 specific antibody (1∶50 dilution) respectively, followed by anti-Rabbit Alexa Fluor 594 and anti-goat Alexa Fluor 555 (red) as secondary antibodies. Endogenous EBNA3C was detected using A10 ascites (1∶150 dilution) followed by anti-mouse Alexa Fluor 488 (green). DAPI (49, 69-diamidino-2-phenylindole) was used (1∶500 dilution) to stain nuclei. The images were captured by Olympus Fluoview confocal microscope.