Journal: Fundamental Research

Article Title: O-GlcNAcylation of YTHDF2 antagonizes ERK-dependent phosphorylation and inhibits lung carcinoma

doi: 10.1016/j.fmre.2024.07.003

Figure Lengend Snippet: Fig. 2 O-GlcNAcylation of YTHDF2 at Thr-49 antagonizes ERK-dependent

Article Snippet: The following primary antibodies were used for immunoblot: anti-Myc (PTM Bio, #PTM-5390, 1:3000), anti-c-Myc (ProteinTech, #10828-1-AP, 1:1000), anti-YTHDF2 (proteinTech, #24744-1-AP, 1:1000), anti-Flag (Sigma, #F1084, 1:1000), RL2 (Abcam, #AB2739, 1:1000), anti-GST (Gene Script, #A00865, 1:1000), anti-HA (Bethyl, #A190-108A, 1:1000), anti-OGT (Santa Cruz Biotechnology, #sc-74546, 1:1000), anti-β-actin (Sigma, #A5441, 1:1000), anti-Ubiquitin (PTM Bio, #PTM-1106RM, 1:1000) , anti-AXIN (ProteinTech,#16541-1-AP, 1:1000) and anti-YTHDF2-pS39 antibodies were generated using the sequence PYLpSPQAR by Dia-An Biotech, Inc. Peroxidase-conjugated secondary antibodies were from JacksonImmuno Research.

Techniques:

Journal: Fundamental Research

Article Title: O-GlcNAcylation of YTHDF2 antagonizes ERK-dependent phosphorylation and inhibits lung carcinoma

doi: 10.1016/j.fmre.2024.07.003

Figure Lengend Snippet: Fig. 3 YTHDF2 O-GlcNAcylation promotes ubiquitination. (a) 293T cells were

Article Snippet: The following primary antibodies were used for immunoblot: anti-Myc (PTM Bio, #PTM-5390, 1:3000), anti-c-Myc (ProteinTech, #10828-1-AP, 1:1000), anti-YTHDF2 (proteinTech, #24744-1-AP, 1:1000), anti-Flag (Sigma, #F1084, 1:1000), RL2 (Abcam, #AB2739, 1:1000), anti-GST (Gene Script, #A00865, 1:1000), anti-HA (Bethyl, #A190-108A, 1:1000), anti-OGT (Santa Cruz Biotechnology, #sc-74546, 1:1000), anti-β-actin (Sigma, #A5441, 1:1000), anti-Ubiquitin (PTM Bio, #PTM-1106RM, 1:1000) , anti-AXIN (ProteinTech,#16541-1-AP, 1:1000) and anti-YTHDF2-pS39 antibodies were generated using the sequence PYLpSPQAR by Dia-An Biotech, Inc. Peroxidase-conjugated secondary antibodies were from JacksonImmuno Research.

Techniques: Ubiquitin Proteomics

Journal: Fundamental Research

Article Title: O-GlcNAcylation of YTHDF2 antagonizes ERK-dependent phosphorylation and inhibits lung carcinoma

doi: 10.1016/j.fmre.2024.07.003

Figure Lengend Snippet: Fig. 4 O-GlcNAcylation of YTHDF2 downregulates c-Myc in H1299 and A549 lung

Article Snippet: The following primary antibodies were used for immunoblot: anti-Myc (PTM Bio, #PTM-5390, 1:3000), anti-c-Myc (ProteinTech, #10828-1-AP, 1:1000), anti-YTHDF2 (proteinTech, #24744-1-AP, 1:1000), anti-Flag (Sigma, #F1084, 1:1000), RL2 (Abcam, #AB2739, 1:1000), anti-GST (Gene Script, #A00865, 1:1000), anti-HA (Bethyl, #A190-108A, 1:1000), anti-OGT (Santa Cruz Biotechnology, #sc-74546, 1:1000), anti-β-actin (Sigma, #A5441, 1:1000), anti-Ubiquitin (PTM Bio, #PTM-1106RM, 1:1000) , anti-AXIN (ProteinTech,#16541-1-AP, 1:1000) and anti-YTHDF2-pS39 antibodies were generated using the sequence PYLpSPQAR by Dia-An Biotech, Inc. Peroxidase-conjugated secondary antibodies were from JacksonImmuno Research.

Techniques:

Journal: Fundamental Research

Article Title: O-GlcNAcylation of YTHDF2 antagonizes ERK-dependent phosphorylation and inhibits lung carcinoma

doi: 10.1016/j.fmre.2024.07.003

Figure Lengend Snippet: Fig. 5 O-GlcNAcylation of YTHDF2 regulates the metastatic capacity of H1299 and

Article Snippet: The following primary antibodies were used for immunoblot: anti-Myc (PTM Bio, #PTM-5390, 1:3000), anti-c-Myc (ProteinTech, #10828-1-AP, 1:1000), anti-YTHDF2 (proteinTech, #24744-1-AP, 1:1000), anti-Flag (Sigma, #F1084, 1:1000), RL2 (Abcam, #AB2739, 1:1000), anti-GST (Gene Script, #A00865, 1:1000), anti-HA (Bethyl, #A190-108A, 1:1000), anti-OGT (Santa Cruz Biotechnology, #sc-74546, 1:1000), anti-β-actin (Sigma, #A5441, 1:1000), anti-Ubiquitin (PTM Bio, #PTM-1106RM, 1:1000) , anti-AXIN (ProteinTech,#16541-1-AP, 1:1000) and anti-YTHDF2-pS39 antibodies were generated using the sequence PYLpSPQAR by Dia-An Biotech, Inc. Peroxidase-conjugated secondary antibodies were from JacksonImmuno Research.

Techniques:

Journal: Fundamental Research

Article Title: O-GlcNAcylation of YTHDF2 antagonizes ERK-dependent phosphorylation and inhibits lung carcinoma

doi: 10.1016/j.fmre.2024.07.003

Figure Lengend Snippet: Fig. 6 YTHDF2 O-GlcNAcylation inhibits lung cancer. (a-c) Xenografts in nude mice.

Article Snippet: The following primary antibodies were used for immunoblot: anti-Myc (PTM Bio, #PTM-5390, 1:3000), anti-c-Myc (ProteinTech, #10828-1-AP, 1:1000), anti-YTHDF2 (proteinTech, #24744-1-AP, 1:1000), anti-Flag (Sigma, #F1084, 1:1000), RL2 (Abcam, #AB2739, 1:1000), anti-GST (Gene Script, #A00865, 1:1000), anti-HA (Bethyl, #A190-108A, 1:1000), anti-OGT (Santa Cruz Biotechnology, #sc-74546, 1:1000), anti-β-actin (Sigma, #A5441, 1:1000), anti-Ubiquitin (PTM Bio, #PTM-1106RM, 1:1000) , anti-AXIN (ProteinTech,#16541-1-AP, 1:1000) and anti-YTHDF2-pS39 antibodies were generated using the sequence PYLpSPQAR by Dia-An Biotech, Inc. Peroxidase-conjugated secondary antibodies were from JacksonImmuno Research.

Techniques:

Journal: Journal of Integrative Agriculture

Article Title: Critical role of cytochrome c1 and its cleavage in porcine reproductive and respiratory syndrome virus nonstructural protein 4-induced cell apoptosis via interaction with nsp4

doi: 10.1016/s2095-3119(17)61670-8

Figure Lengend Snippet: Fig. 1 Porcine reproductive and respiratory syndrome virus (PRRSV) nonstructural protein 4 (nsp4) interacts with cytochrome c1 (cyto.c1). A, yeast two hybrid assay. The plasmids pGBKT7-nsp4 and pGADT7-cyto.c1 were co-transformed into the yeast strain Y2HGold and selected on QDO/X/ABA plates. The top and second panels are the respective positive and negative controls. B, HEK 293T cells were transfected to express Flag-cyto.c1, or HA-nsp4 or together. At 24 h post-transfection, the cells were either subjected to direct Western blot analysis or lysed and immunoprecipitated with anti-HA antibodies. The proteins bound to sepharose beads were separated by SDS-PAGE, transferred to PVDF membranes, and probed with the antibodies to Flag and HA. C, the same as B except that nsp4 truncation mutants were used. D, MARC-145 cells were transduced with letiviruses that were expressing GFP or nsp4-GFP in the presence of 8 µg mL–1 polybrene, respectively. The cells were harvested 48 h post-transduction and cell lysates were immunoprecipitated using anti-GFP beads. The proteins bound to sepharose beads were subjected to Western blot analysis using an anti-cytochrome c1 polyclonal antibody or anti-GFP mAb. E, PRRSV nsp4 interacts with cyto.c1 in mammalian cells. Vero cells grown on coverslips in six-well plates were transfected when 60–70% to express the indicated proteins, either alone or pairwise. At 18–24 h post-transfection, the cells were fixed, permeablized, and stained with antibodies to HA and Flag, and examined by confocal microscopy. For double transfections, the merged images are shown at the right.

Article Snippet: Mouse anti-green fluorescent protein (GFP) mAb (66002-1-Ig) and rabbit anti-cytochrome c1 polyclonal antibodies (10242-1-AP) were purchased from Proteintech (Chicago, IL, USA).

Techniques: Virus, Y2H Assay, Transformation Assay, Transfection, Western Blot, Immunoprecipitation, SDS Page, Transduction, Expressing, Staining, Confocal Microscopy