Immunofluorescent analysis of mCherry Tag was performed using 70% confluent construct name transfected HEK-293 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with mCherry Polyclonal Antibody (Product # PA5-34974) at 1:100 dilution and Histone H3 Monoclonal Antibody (865R2) (Product # AHO1432) at 1:200 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 647 (Product # A32733) and Goat anti-Mouse IgM (Heavy chain) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 (Product # A21042) respectively at a dilution of 1:2,000 for 45 minutes at room temperature. Panel a (Nuclei: Red) represents mCherry Tag. Panel b (Nuclei: Green) represents Histone H3. Panel c (Nuclei: Blue) represents ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). Panel d represents the merged image showing the co-localization of nuclear signals in transfected cells. Panel e represents untransfected HEK-293 cells. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.