For western blot analysis, antibodies directed against the following proteins were used: GFP (SC-9996, Santa Cruz, 1:2000), α-tubulin (T6074, Sigma, 1:10000), α-tubulin (Ab18251, Abcam, 1:10000), GAPDH (60004-1lg, Proteintech, 1:20000), HA (16B12, Covance, 1:1000), GIGYF2 (NBP2-12812, Novusbio, 1:3000), FLAG-tag (F1804, Sigma, 1:500), DDX6 (A300-461A, Bethyl Laboratories, 1:1000) and GIGYF1 (A304-132A-M, Bethyl Laboratories, 1:500). Secondary antibodies were: IRDye 800CW goat anti-rabbit (LICOR, 1:10000), IRDye 680RD goat anti-rabbit (LICOR, 1:10000), IRDye 800CW goat anti-mouse (LICOR, 1:10000) and Alexa 680 goat anti-mouse (A21057, Life Technologies 1:10000). For Immunoprecipitation: HA (Covance), GIGYF2 (Novusbio) and mouse or rabbit IgG (Sigma). For Immunofluorescence: GIGYF2 (NBP2-12812, Novusbio, 1:500), HA (16B12, Covance, 1:1000), anti-mouse Alexa 488 (A28175, Invitrogen, 1:1000) and anti-rabbit Alexa 633 (A21070, Invitrogen, 1:1000).
Amaya Ramirez C.C., Hubbe P., Mandel N, & Béthune J. (2018). 4EHP-independent repression of endogenous mRNAs by the RNA-binding protein GIGYF2. Nucleic Acids Research, 46(11), 5792-5808.
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