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. 2019 Jan 18;9:208. doi: 10.1038/s41598-018-36312-4

Figure 5.

Figure 5

Increased SUMO levels were detected in LMP1-positive lymphoma tissues and cells. (a) FFPET biopsy tissues were sectioned onto glass slides. Following staining with rabbit anti-SUMO-1-specific and mouse anti-LMP1-specific primary antibodies and goat anti-mouse Alexa Fluor 488 or goat anti-rabbit Alexa Fluor 594 secondary antibodies, coverslips were mounted using ProLong® Gold Antifade Reagent with DAPI. Immunofluorescence microscopy was performed at 20X magnification and images created using the Openlab software. (b) Serum-starved 293 cells were transfected with FLAG-LMP1-expression constructs. 24 hours post-transfection, cells were fixed, permeabilized, and stained with rabbit anti-SUMO-1-specific and mouse anti-FLAG-specific primary antibodies and goat anti-mouse Alexa Fluor 488 or goat anti-rabbit Alexa Fluor 594 secondary antibodies. Coverslips were mounted using ProLong® Gold Antifade Reagent with DAPI. Confocal microscopy was performed at 60X magnification using the Nikon A1 laser confocal microscope. Images of LMP1-transfected cells next to non-LMP1-expressing cells were captured. Representative images are shown. (c) The corrected total cell fluorescence was determined for SUMO-1 levels in approximately 150 LMP1-negative and 150 LMP1-positive cells in lymphoma tissue samples and approximately 65 LMP1-negative and 65 LMP1-positive 293 cells. Results are shown as the mean ± the standard deviation.