Journal: Journal of Translational Medicine
Article Title: LDHC4 drives lung adenocarcinoma progression by inducing lactylation of RB1 at lysine 900 to disrupt the RB1–E2F1 complex
doi: 10.1186/s12967-026-08070-9
Figure Lengend Snippet: Influence of RB1–K900lac on the cell cycle pathway. ( A to C ) Cell cycle analysis by flow cytometry in A549 and PC-9 cell lines stably expressing RB1–WT or RB1–K900R via lentiviral vectors. ( D ) Representative immunofluorescence images showing the distribution of CDK1 in A549 and PC-9 cells. CDK1 protein was labeled with red fluorescent Cy3, and nuclei were counterstained with blue fluorescent DAPI. Images were acquired using a high-resolution confocal multiphoton microscopy system (NIKON AX RMP, Japan). ( E , F ) Western blot analysis of cell cycle-related CDK molecule expression. ( G , H ) Expression of cell cycle-related cyclin molecules. ( I , J ) Expression of P21 and Chk1 molecules. ( K ) Schematic diagram illustrating how LDHC4 promotes the cell cycle by inducing RB1 lactylation. ** p < 0.01, *** p < 0.001
Article Snippet: The following antibodies were used in this study: rabbit anti-human LDHC (subunit C) monoclonal antibody (mAb) (Proteintech Group, Inc., 1:1000), rabbit anti-human RB1 mAb (Proteintech Group, Inc., 1:2000), rabbit anti-human L-Lactyl Lysine mAb (PTM Bio, Inc., 1:1000), rabbit anti-human E2F1 mAb (APExBIO Technology, LLC, 1:500), rabbit anti-human Lamin B mAb (Beyotime Biotech, Inc., 1:1000), rabbit anti-human CDK1 mAb (Beyotime Biotech, Inc., 1:800), rabbit anti-human CDK2 mAb (Beyotime Biotech, Inc., 1:800), rabbit anti-human CDK4 mAb (Beyotime Biotech, Inc., 1:1000), rabbit anti-human CDK6 mAb (Beyotime Biotech, Inc., 1:1000), rabbit anti-human cyclin A2 mAb (Beyotime Biotech, Inc., 1:1000), rabbit anti-human cyclin B1 mAb (Beyotime Biotech, Inc., 1:1000), rabbit anti-human cyclin D1 mAb (Beyotime Biotech, Inc., 1:500), rabbit anti-human P21 mAb (Abmart Bio, Inc., 1:500), rabbit anti-human Chk1 mAb (Immunoway Bio, Inc., 1:2000), and rabbit anti-β-Actin monoclonal antibody (Beyotime Biotech, Inc., 1:1000). β-Actin and Lamin B served as loading controls, and protein band intensities were quantified using Image J software.
Techniques: Cell Cycle Assay, Flow Cytometry, Stable Transfection, Expressing, Immunofluorescence, Labeling, Microscopy, Western Blot