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

Journal: Cell Death & Disease

Article Title: The ATC12 small molecule inhibits the Aurora-A/TPX2 interaction and impairs the proliferation of breast cancer cells

doi: 10.1038/s41419-026-08579-3

Figure Lengend Snippet: A Representative images of time-lapse video recording of MDA-MB-231 cells treated with DMSO, 30 or 50 μM ATC12 for 72 h, acquired at t0, 24, 48, and 72 h. The percentage of dividing cells within 24 h intervals is shown in ( B ), while the time spent in mitosis (minutes from the round-up to the complete re-attachment of daughter cells) is shown in ( C ). The graph in ( D ) indicates the increase (F.C., fold change) in cells at 24, 48, and 72 h, with respect to the initial number of cells (t0) per condition. Sample size per condition, from two (ATC12 30 μM) or three (DMSO and ATC12 50 μM) independent experiments: DMSO: 231 ( B , D ), 356 ( C ); ATC12 30 μM: 255 ( B , D ), 261 ( C ); ATC12 50 μM: 221 ( B , D ), 190 ( C ). E Representative IF images of p21 staining in MDA-MB-231 cells treated with 50 μM ATC12 for 72 h. The graphs show: quantification of p21 nuclear signal intensity (left, DMSO: 1051 cells; ATC12: 415 cells, from three independent experiments) and the nuclear size (right, DMSO: 829 cells; ATC12: 587 cells, from three independent experiments). F Representative panels of C 12 FDG staining in MDA-MB-231 cells treated with 50 μM ATC12 for 72 h. C 12 FDG signal intensity per field (DMSO: 1919 cells; ATC12: 1058 cells, from three independent experiments) is quantified in the graph. The control condition is set as 1 in relative quantifications. Error bars: SD; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001; Fisher’s exact test ( B ), Kruskal–Wallis test ( C ), Two-way ANOVA multiple comparisons test ( D ), Mann–Whitney test ( E , F ). Scale bars, 20 μm ( A ); 10 μm ( E , F ).

Article Snippet: Primary antibodies were as follows: rabbit anti-phospho-Aurora-A (Thr288; #3079S, 1:100, Cell Signaling Technology, Danvers, Massachusetts, USA); mouse anti-α-tubulin (T5168, 2 μg/ml, Sigma Aldrich-Merck KGaA, Darmstadt, Germany); mouse anti-Aurora-A (610939, 0.5 μg/ml, BD Transduction Laboratories, Meylan, France); rabbit anti-TPX2 (NB500-179, 1:1500, Novus Biologicals, Cambridge, UK); rabbit anti-p21 (2947, 1:1000, Cell Signaling Technology, Danvers, Massachusetts, USA).

Techniques: Staining, Control, MANN-WHITNEY

Journal: Cellular Oncology

Article Title: Targeting senescence-like tumor-associated macrophages sensitizes chemotherapy in triple-negative breast cancer

doi: 10.1007/s13402-026-01197-3

Figure Lengend Snippet: Senescence-like TAMs are induced upon chemotherapy in vivo. (A) CD163⁺ TAMs were isolated from human breast cancer tissue obtained via needle biopsy (pre-treatment) and surgical resection (post-neoadjuvant chemotherapy). Isolated TAMs underwent proteomic profiling using LC-MS/MS. The dataset is downloaded from the PRIDE repository under accession PXD022673. (B) Gene Set Enrichment Analysis (GSEA) of proteomic data highlights significantly altered biological functions in TAMs following neoadjuvant chemotherapy. (C) GSEA reveals significant upregulation of cellular senescence signatures in TAMs isolated from ADM-treated orthotopic 4T1 tumors, compared to PBS-treated controls. The dataset is downloaded from the PRIDE repository under accession PXD022674. (D) IF analysis of p16 + F4/80 + and p21 + F4/80 + TAMs isolated from treatment-naïve and ADM-treated tumors ( n = 5). Scale bar, 50 μm. (E) Real-time qPCR analysis of senescence-related genes ( Cdkn2a , Cdkn1a , and Trp53 ) in TAMs isolated from treatment-naïve and ADM-treated tumors ( n = 5). (F) Western blotting analysis of senescence markers (p16 and p21) in TAMs isolated from treatment-naïve and ADM-treated tumors ( n = 5). (G) ELISA quantification of senescence-associated secretory phenotype (SASP) markers (IL-1α, IL-6, CXCL1, TNF-α) in conditioned media from TAMs isolated from orthotopic 4T1 tumors treated with control or ADM ( n = 5). For statistical analyses, an unpaired two-tailed t-test was used. * P < 0.05, ** P < 0.01

Article Snippet: The primary antibodies employed were: F4/80 (Cell Signaling Technology, Cat. 30325; RRID: AB_2798990), CD163 (Abcam, Cat. 156769; RRID: AB_3076143), p21 (Cell Signaling Technology, Cat. 2947; RRID: AB_823586), and p16 (Abcam, Cat. 211542; RRID: AB_2891084).

Techniques: In Vivo, Isolation, Liquid Chromatography with Mass Spectroscopy, Western Blot, Enzyme-linked Immunosorbent Assay, Control, Two Tailed Test