Prolong gold antifade mountant with dapi for nuclear staining

After the deparaffination and rehydration process, antigens were unmasked by steaming in Antigen Retrieval Citra, pH-6.0 (#HK086-9K, BioGenes, Cupertino, CA, USA) for 10 min. After washing in PBS, sections were incubated in 0.1 mg/mL BODIPY^TM 492/515 (#D3922, Invitrogen) in DMSO for 30 min at room temperature. Sections were washed in PBS and mounted with ProLong^TM Gold Antifade Mountant with DAPI for nuclear staining (#P36931, Invitrogen). All stained sections were viewed with a fluorescence microscope (Revolve, Echo, San Diego, CA, USA).

After the deparaffination and rehydration process, tissue sections were steamed in Antigen Retrieval Citra, pH-6.0 (#HK086-9K, BioGenex, Fremont, CA, USA) for 20 min. After several washes, PowerBlock (#HK085-5K, BioGenex, Fremont, CA, USA) was applied to each section according to the product directions. The sections were then incubated in the anti-FoxO1 (#2880, Cell Signaling) in 1:100 ratio at 4 °C overnight. After several washes, sections were incubated in the anti-rabbit secondary antibody (#A11008, Invitrogen) in 1:20 ratio at room temperature for one hour. Sections were then washed with PBS and mounted with ProLong^TM Gold Antifade Mountant with DAPI for nuclear staining (#P36931, Invitrogen). All stained sections were viewed with a fluorescence microscope (Revolve, Echo, San Diego, CA, USA).