Immunocytochemistry analysis of ATF5 in RAW264.7 cells. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with ATF5 monoclonal antibody (Product # MA5-32365) using a dilution of 1:100 in 1% BSA in PBST overnight at 4°C followed by Goat Anti-Rabbit IgG H&L (iFluor™ 488) secondary antibody at a dilution of 1:1,000. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1:100 dilution overnight at 4°C. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1:1,000 dilution.