Immunofluorescence analysis of XRN1 was performed using SH-SY5Y and SH-SY5Y treated with BDNF (100 ng/mL, 72 hours). 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 XRN1 Polyclonal Antibody (Product # PA5-57110) at 5 µg/mL concentration in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, Alexa Fluor 488 (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 Rhodamine Phalloidin (Product # R415, 1:300). Panel d and e represents the merged image showing cytoplasmic localization. Expression of XRN1 was increased upon BDNF treatment in SH-SY5Y cells (Panel d) as compared to untreated SH-SY5Y cells (Panel e). Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.