Immunofluorescence analysis of GOT1 was performed using 70% confluent log phase Hep G2 cells treated with Insulin (100 nanomolar for 8h). The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with GOT1 Polyclonal Antibody (Product # PA5-18845) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Rabbit anti-Goat IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 (Product # A-11078) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing decrease in expression of GOT1 in cytoplasm upon Insulin treatment. Panel e represents untreated cells and panel f shows control cells with no primary antibody to assess background. The images were captured at 60X magnification.