Immunofluorescence analysis of RNPS1 was performed using 70% confluent log phase HeLa cells. 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 RNPS1 Polyclonal Antibody (Product # PA5-35967) at 1:200 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing predominant nuclear localization along with weak cytoplasmic signal. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Western blot analysis was performed on modified whole cell extracts (1% SDS) (30 µg lysate) of A-431 (Lane 1), HeLa (Lane 2), Hep G2 (Lane 3), and Raji (Lane 4). The blot was probed with Anti-RNPS1 Polyclonal Antibody (Product # PA5-35967, 1:2000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 48 kDa band corresponding to RNPS1 was observed in all cell lines tested along with an additional uncharacterized band (*) at ~40 kDa.