Anti flag m20008

Zebrafish wild type AB line was purchased from the China zebrafish resource center. MiR-202-5p^-/- zebrafish were generated in our laboratory previously [17 (link)]. Fish were raised at 28 °C with 10 h darkness and 14 h light and were fed with commercial pellets twice a day. All embryos were collected after natural spawning and staged as previously reported [18 (link)].
ZBE3 cells derived from zebrafish embryos were cultured at 28 °C as previously described [19 (link)]. HEK 293T cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with heat inactivated 10% FBS (Invitrogen, California, USA), pyruvate glucose, and L-glutamine (Gibco, California, USA), at 37 °C under a humidified atmosphere of air containing 5% CO₂.
The anti-Flag (M20008) and anti-HA antibodies (M20013) were purchased from Abmart (Guangzhou, China). Anti-α-Tubulin (ab15246) and anti-GFP antibodies (G1544) were purchased from Abcam (London, Britain) and Sigma (Missouri, USA), respectively. The secondary antibodies goat anti-rabbit IgG-HRP and goat anti-mouse IgG-HRP were purchased from Invitrogen.
miRNA mimics for miR-202-5p, hsa-miR-202-5p, and negative controls of miRNA mimics (Ctrl-m) were purchased from Ribobio (Guangzhou, China).

Anti-Moesin (ab151542) (1:1000), anti-Ezrin (ab4069) (1:200), anti-Radixin (ab52495) (1:1000), anti-phospho-ERM (ab76247) (1:1000), and anti-CD8α (ab209775) were purchased from Abcam. anti-Zap-70 (2705) (1:1000), anti-Syk (2712) (1:1000), anti-phospho-Erk1/2 (8544, 9101) (1:1000), anti-Erk1/2 (9102) (1:1000), anti-phospho-Akt (9271) (1:800), anti-Akt (9272) (1:1000), anti-phospho-P38 (9211) (1:1000), anti-P38 (9212) (1:1000), anti-rabbit-IgG (7074), and anti-mouse-IgG (7076) were purchased from CST. Anti-phospho-tyrosine (05–321) (1:1000) was purchased from Merck. Anti-GAPDH (BE0023) (1:3000) was purchased from EASYBIO. Anti-Myc (M20002) (1:1000), anti-Flag (M20008) (1:1000), and Protein A/G-Agarose (A10001) were purchased from Abmart. Anti-Flag M2 affinity gel (A2220) and uric acid (U2625) were purchased from Sigma-Aldrich. Polystyrene microspheres (19822) were purchased from Polysciences. Piceatannol (527948), Syk Inhibitor (574711), Syk Inhibitor IV, BAY 61-3606 HCL (574714), R406 (5.05819.0001), Wortmannin (681676), and LY294002 (440204) were purchased from Calbiochem. AZD0530 (S1006) and PP2 (S7008) were purchased from Selleck. Bodipy FL-PI(4,5)P2 (C-45F6) were purchased from Echelon Biosciences. TopFluor TMR-PC (810180C) and TopFluor TMR-PE (810241C) were purchased from Avanti Lipids. Geneticin (10131027) was purchased from ThermoFisher.

Huh7, HepG2, and HEK293T cells were cultured in Dulbecco’s modified Eagle medium (Bio-Channel, China) supplemented with 10% fetal bovine serum (Bio-Channel, China) and penicillin-streptomycin. Flag-tagged RIG-I, MDA5, MAVS, TBK1, IRF3, HA-tagged WT-Ub, K48-Ub, and K63-Ub plasmids were kept in our laboratory. HA-tagged USP15 were created by cloning the CDS (coding sequence) region of USP15 into the pcDNA3.1-HA vector. Plasmids, miRNA mimics, and inhibitors were transfected into cells using M5 Hiper Lip.2000 (Mei5bio) according to the manufacturer’s instructions. All the antibodies used in this research were listed as follows: anti-Flag (M20008) and anti-HA (M20003) antibodies were purchased from Abmart; anti-RIG-I (20566–1-AP), anti-USP15 (14354–1-AP), anti-Ub (10201–2-AP), and anti-GAPDH (60004–1-Ig) were purchased from Proteintech; anti-K48-Ub (8081), anti-K63-Ub (5621), and anti-Phospho-IRF3 (37829) were obtained from Cell Signaling Technology.