Journal: Cell Communication and Signaling : CCS

Article Title: LDOC1 connects histone H2B monoubiquitination to tumor cell plasticity in non-small cell lung cancer

doi: 10.1186/s12964-025-02607-z

Figure Lengend Snippet: LDOC1 loss alters TGF-β–driven EMT programs and motility through H2Bub1 dysregulation. a Confocal images of A549 sublines treated with TGF-β (10 ng/mL) for the indicated days and stained with FITC–anti-α-tubulin (DM1A) and DAPI. Scale bars, 20 μm. b Time-course qRT–PCR of EMT- and cytoskeleton-related genes in A549-shCtrl-1 and A549-shLDOC1-1 cells after TGF-β stimulation; expression was normalized to GAPDH (mean ± SEM, n = 3). c–e Transwell migration of A549-shCtrl-1/-2 and A549-shLDOC1-1/-2 cells (c) and H1299 cells transfected with vector (V) or LDOC1 ( d) , with quantitative summary in ( e ) (mean ± SEM, n = 3). f–h Matrigel invasion assays for the same A549 (f) and H1299 (g) panels, with quantification in (h) (mean ± SEM, n = 3). i Incucyte wound-healing assays showing wound confluence over time in A549-shCtrl and A549-shLDOC1 sublines with or without TGF-β; representative images at 0 h and 48 h are shown. j Immunoblot of H2Bub1 in A549-shLDOC1-1 cells expressing H2BK120R mutants (clones B10 and C9); Lamin B1, loading control; V, vector. k–m Matrigel invasion ( k ) with quantification ( l ) and wound-healing analysis (m) of A549-shLDOC1-1 cells expressing V or H2BK120R mutants (B10, C9) (mean ± SEM, n = 3). * P < 0.05, ** P < 0.01 (two-tailed t-test)

Article Snippet: Chemical reagents included recombinant human TGF-β (MCE, HY-P7118), the proteasome inhibitor bortezomib (MCE, HY-10227) and recombinant human EGF (MCE, HY-P7109).

Techniques: Staining, Quantitative RT-PCR, Expressing, Migration, Transfection, Plasmid Preparation, Western Blot, Clone Assay, Control, Two Tailed Test

Journal: Cell Communication and Signaling : CCS

Article Title: LDOC1 connects histone H2B monoubiquitination to tumor cell plasticity in non-small cell lung cancer

doi: 10.1186/s12964-025-02607-z

Figure Lengend Snippet: LDOC1 loss promotes a hybrid epithelial–mesenchymal state and elevated H2Bub1 associated with STAS and clinical outcome in NSCLC. a Immunoblot analysis of LDOC1 and EMT markers in A549-shCtrl-1/-2 and A549-shLDOC1-1/-2 cells; GAPDH and β-actin served as loading controls. b Confocal immunofluorescent images of E-cadherin (green) and nuclei (DAPI, blue) in A549 sublines. Scale bars, 10 μm. c–e Flow-cytometric analysis of A549-shCtrl-1 and A549-shLDOC1-1 cells stained for E-cadherin–APC and vimentin–PE: (c) representative single-parameter histograms, (d) bivariate plots showing E-cad⁺/Vim⁻, E-cad⁻/Vim⁺ and E-cad⁺/Vim⁺ populations, and (e) quantification of hybrid E/M (E-cad⁺/Vim⁺) cells (mean ± SEM; *P < 0.05, two-tailed t-test). f Adhesion of A549-shCtrl-1/-2 and A549-shLDOC1-1/-2 cells with or without TGF-β (mean ± SEM, n = 3; *P < 0.05, **P < 0.01, ***P < 0.001, two-tailed t-test). g–h Representative IHC for H&E, E-cadherin and vimentin (g) and H2Bub1 (h) in paired primary tumors and matched STAS lesions from NSCLC; insets show higher magnification. Scale bars, 100 μm. i Kaplan–Meier curves of progression-free survival in chemotherapy-treated NSCLC patients stratified by H2Bub1 expression. j LDOC1 mRNA levels in KRAS-mutant versus KRAS^WT tumors in the TCGA LUAD cohort (n = 520; Welch’s t-test)

Article Snippet: Chemical reagents included recombinant human TGF-β (MCE, HY-P7118), the proteasome inhibitor bortezomib (MCE, HY-10227) and recombinant human EGF (MCE, HY-P7109).

Techniques: Western Blot, Staining, Two Tailed Test, Expressing, Mutagenesis