Immunofluorescent Localization of IGF2BP2 and IGF2BP3

Immunofluorescence staining was performed to detect the expression of IGF2BP2 and IGF2BP3. In brief, the tissue samples were deparaffinization, blocked at room temperature for 60 min with a blocking solution (5% BSA/0.1% Triton X-100 PBS), and stained with the primary anti-IGF2BP2 (1:200; 11601-1-AP; Proteintech) and anti-IGF2BP3 (1:200; 14642-1-AP; Proteintech) antibodies. The samples were then washed with Tris-buffered saline for 30 min, and incubated with the secondary goat anti-rabbit IgG H&L (Alexa Fluor^® 568) (1:200, ab175471; Abcam) and goat anti-mouse IgG H&L (Alexa Fluor 488) (1:200, ab150113; Abcam) antibodies for 30 min at 37 °C in the dark. After washing in PBS, DAPI staining was performed for 5 min, to counterstain the nuclei (28 (link)). Finally, the samples were sealed in 50% glycerin and imaged on a fluorescence microscope (DP72, OLYMPUS, Japan).