3d). In addition to the crucial role of CD8 T cell immunity, we were also curious about whether CD4 T cells were affected in Ythdf2CKO mice. However, antibody-mediated neutralization confirmed that YTHDF2 loss mainly jeopardizes CD8 (but not CD4) T cell-mediated antitumor immunity (...
66,67] and was remarkably downregulated in clear cell renal cell carcinoma (ccRCC) [68]. A lack of YTHDF2 could significantly decrease the migration rate and reduce the expression level of related proteins in bladder cancer cells, indicating that YTHDF2 acts as an oncogene in...
Additionally, DDIT4 mRNA precipitated by the m6A antibody decreased markedly in Mettl3-knockdown CAL-62 cells (Fig. 4H). Subsequently, the effects of YTHDF2 knockdown and overexpression on the stability of DDIT4 mRNA was studied. The stability of DDIT4 mRNA was increased in CAL-62 cells ...
Next, 50 µL of antibody was added to each well, followed by the addition of a fluorescent solution. The plates were then incubated at room temperature in the dark for 4 min. The absorbance was subsequently measured at 450 nm using an enzyme marker. Cell cultures and transfection Initially...
The membranes were blocked with 5% non-fat milk for 1 h at room temperature and incubated with primary antibodies and HRP-conjugated secondary antibody. The chemiluminescent signaling was detected by using ECL reagents (Pierce, USA). Primary antibodies used in this study: anti-YTHDF2 (24744-...
Ceramide and S1P levels in cell extracts were determined by ELISA kits (mlbio, Shanghai, China), respectively. Cellular ceramide or S1P was captured by solid-phase antibody and then combined with horseradish peroxidase (HRP)-labeled detection antibody to form an antibody-antigen-enzyme-antibody comp...
signaling. Selectively targeting YTHDF2 in TAMs using a Toll-like receptor 9 agonist-conjugated small interfering RNA reprogrammed TAMs toward an antitumoral phenotype, restrained tumor growth and enhanced the efficacy of PD-L1 antibody therapy. Collectively, our findings describe the role of YTHDF2...
The supernatant was incubated with antibodies against YTHDF2 antibody (Abcam, USA) overnight at 4 °C. Then, 50 μl A/G magnetic beads were added to the supernatant and incubated for 6 h. After immobilizing the magnetic bead bound complexes with a magnetic separator (Millipore USA), ...
This may be because YTHDF2 downregulation results in decreased cytotoxic CD8+ T cell infiltration, suppressing the efficacy of PD-1 blockade. Then, the combination of OXA and anti-PD1 antibody were performed after specifically overexpressing CX3CL1 in hepatocytes using the AAV (Supplementary Fig. ...
After lysis, the cell extracts were centrifuged at 14,000 g for 15 min to separate the supernatant. The cleared lysate was then incubated with magnetic bead protein A/G (Thermo Fisher, #88802), which had been pre-conjugated with either 5 µg of control IgG antibody (Thermo Fisher, #...