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To investigate whether the recruitment of TRIM25 into the SGs is specific to poly(I:C) treatment, we systematically treated HeLa cells that overexpressed GFP-TRIM25 with various types of stress inducers: RNA virus invasion (Sendai virus, SeV), exogenous dsRNA stress (poly(I:C), oxidative stre...
Immunoblotting was performed using MBP antibody (AE016;ABclonal;1:5000 dilution) and His antibody (TA150088;ORIGENE;1:5000 dilution) to detect the presence of the respective proteins. For histone peptide pull-down assays, 2 μg of GST-fused protein was incubated with 1 μg of...
DUSP18, USF1 and their corresponding mutations were subcloned into the pHAGE-CMV-MCS-PGK-3×Flag, pCMV-HA, or pLKO.1-GFP empty vector, the PCR primer sequence used in this study is summarized in Supplementary Table 2. The lentivirus vectors were co-transfected with psPAX2 plasmid and ...
Xuemei Yan, Jiayun Liu, Ruizhen Li, Siqi Ge, Sai Wang, Xingan Liu,et al.Solarbio, K200060M), anti-GFP(1:3000dilution; Solarbio, K114305P), anti-Flag (1:5000dilution;CST, #14793), anti-Actin (1:2000dilution; Biopm, PMK085S), anti-HA...
For shRNA experiments, shScrambled vector and shFGF2 vector were transfected into 5–8F cell line with GFP using a lentiviral system (GeneCopoeia Inc., USA). Briefly, 293T cells were transfected with lentiviral vectors for lentivirus production. Cells were cultured for an additional 72 h, ...
An in vivo liver-stage parasite inhibition assay was conducted as described previously68,69. Briefly, five female BALB/c mice (6 weeks, 18 g) per group were infected with 5000 P. berghei-ANKA-GFP-lucsporozoites by i.v injection followed immediately by i.v.-administered JX21108 at a...
For generating CRISPR-mediated knockout cell lines, sgRNAs were sub-cloned into pLenti-CRISPRV2 GFP vector (Addgene). The sgRNA sequences, for human AMPKα1: sg1- 5'GAAGATCGGCCACTACATTC3', sg2-5'TACATTCTGGGTGACACGCT3'; for human AMPKα2: sg1-5'TCAGCCATCTTCGGCGCGCG3', sg2-5'GAAGA...
β-Catenin and GFP mark stem cells. c Intestinal tumor number in the indicated genotypes (n = 15 mice per group; ***p < 0.001, t-test; repeated three independent times). d Percent survival curves of mice with the indicated genotypes (n = 15 mice per group; ***p <...
(D–G) GFP-labeled B16 tumor cells (n = 3 per group of scramble, LSD1 KO, and LSD1/MDA5 DKO) were isolated from tumor-bearing immunocompetent mice and subjected to RNA-seq analysis. Differential gene expression was shown in volcano plots (D). Dots in red represent increased genes (lo...