Immunofluorescence analysis of Adiponectin was performed using 70% confluent 3T3-L1 cells differentiated with StemPro Adipogenesis Supplement (Product # A10070-01) for 5 days. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Adiponectin Rabbit Polyclonal Antibody (Product # PA1-054) at 2 µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conj µgate (Product # A27034) 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 Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e is undifferentiated cells with no signal. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.