Journal: bioRxiv

Article Title: RNA polymerase loss by nuclear rupture drives LMNA cardiomyopathy

doi: 10.64898/2026.04.03.716433

Figure Lengend Snippet: A) Immunofluorescence for bromouridine (BrU) after cardiomyocytes were cultured with BrU for 2 hr to label nascent RNAs. One binucleated cardiomyocyte per image, with nuclei zoomed in. Texts and icons below images indicate nuclear states. Scale bar: 5 μm. All data in , except , are from cardiomyocytes freshly isolated from mice at day 14 post tamoxifen. B) Nuclear BrU intensity by nuclear states after 1-hr BrU incubation. Density and box plots: BrU intensity distribution and interquartile range. Circles: mean intensity within biological replicates (color coded). Asterisks: P < 0.05 from t -tests on linear regression-estimated means with mouse-clustered standard errors. Underlying data: 244 intact nuclei from 4 WT mice, 275 intact, 111 ruptured, 104 resealed nuclei from 3 Lmna CKO mice. C) Relationship between nuclear BrU intensity and nuclear NLS–tdTomato intensity in all types of nuclei in Lmna CKO cardiomyocytes (519 nuclei from 3 mice). Line: simple linear regression fit with 95% confidence interval. R: Pearson correlation coefficient. P: t -test p -value on linear regression-estimated means with mouse-clustered standard errors. See for individual replicates. D) Immunofluorescence for RNA polymerase II (Pol II) in cardiomyocytes. Scale bar: 5 μm. E) Nuclear Pol II intensity by nuclear states. Underlying data: 285 intact nuclei from 3 WT mice, 180 intact, 133 ruptured, 136 resealed nuclei from 3 Lmna CKO mice. Graph annotations and statistics as in (B) . F) Relationship between Pol II intensity and NLS–tdTomato intensity in all types of nuclei in Lmna CKO cardiomyocytes (495 nuclei from 3 mice). Graph annotations and statistics as in (C) . G) Pol II ChIP-seq and input read coverage in WT and Lmna CKO cardiomyocytes (3 mice per genotype). ChIP-seq signals are normalized to internal spike-in controls. H) Average Pol II ChIP-seq signals across 21,177 protein-coding genes. X-axis: 100 equally-spaced bins in gene bodies, 5 bins for 1 kb-upstream regions, and 10 bins for 2 kb-downstream regions. I) Statistical comparison of gene-body Pol II signals in Lmna CKO versus WT cardiomyocytes for 11,942 Pol II-bound genes. Pol II-lost or gained genes are defined at limma p -value < 0.05. J) Ten most enriched Gene Ontology terms among the 1,759 Pol II-lost genes in Lmna CKO cardiomyocytes, with three representative genes for each term. P: Metascape p -value. K) Gene expression state of Pol II-lost, Pol II-gained, and all other genes in Lmna CKO (n=5) versus WT (n=7) hearts. P, DESeq2 p -value. RNA-seq data from En et al. 2024. L) Summary of . Nuclear rupture causes transcriptional deficiency due to RNA Pol II loss.

Article Snippet: Immunofluorescence images of human myocardial sections were acquired using a Yokogawa CSU-W1 Sora spinning disk confocal microscope with 40x or 60x objectives (40x for normal hearts and 60x for ACTC1-variant heart and LMNA-R541C heart).

Techniques: Immunofluorescence, Cell Culture, Isolation, Incubation, ChIP-sequencing, Comparison, Gene Expression, RNA Sequencing

Journal: bioRxiv

Article Title: RNA polymerase loss by nuclear rupture drives LMNA cardiomyopathy

doi: 10.64898/2026.04.03.716433

Figure Lengend Snippet: A) Relationship between nuclear BrU intensity and nuclear NLS–tdTomato intensity in cardiomyocytes for each of 3 Lmna CKO mice. Line: simple linear regression fit with 95% confidence interval. R: Pearson’s correlation coefficient. All data in are derived from mice at day 14 post tamoxifen. B) Relationship between nuclear RNA Pol II intensity and nuclear NLS-tdTomato intensity in cardiomyocytes for each of 3 Lmna CKO mice. Other details as in (A) . C) Immunofluorescence for Pol II CTD phospho-Ser5 (Pol II Ser5p) in isolated cardiomyocytes expressing NLS-tdTomato and GFP-icGAS. Scale bar: 5 μm. D) Nuclear Pol II Ser5p intensity by nuclear states. Density and box plots (interquartile range): signal distribution of all nuclei. Circles: mean intensity within individual biological replicates (color coded). Asterisks: P < 0.05 from t-tests on linear regression-estimated means with mouse-clustered standard errors. Underlying data: 218 intact nuclei from 3 WT mice, 187 intact, 111 ruptured, 91 resealed nuclei from 3 Lmna CKO mice. E, F) Relationship between Pol II Ser5p intensity and NLS–tdTomato intensity in nuclei of Lmna CKO cardiomyocytes. (E) 424 nuclei from three biological replicates (mice). (F) Analysis within individual biological replicates. Other details as in (A) . G) Immunofluorescence for Pol II CTD phospho-Ser2 (Pol II Ser2p) in isolated cardiomyocytes expressing NLS-tdTomato and GFP-icGAS. Scale bar: 5 μm. H) Nuclear Pol II Ser2p intensity by nuclear states. Underlying data: 198 intact nuclei from 3 WT mice, 233 intact, 101 ruptured, 131 resealed nuclei from 3 Lmna CKO mice. N.S.: not significant. Other details as in (D) . I, J) Relationship between Pol II Ser2p intensity and NLS-tdTomato intensity in nuclei of Lmna CKO cardiomyocytes. (I) 517 nuclei from three biological replicates (mice). (J) Analysis within individual biological replicates. Other details as in (A) .

Article Snippet: Immunofluorescence images of human myocardial sections were acquired using a Yokogawa CSU-W1 Sora spinning disk confocal microscope with 40x or 60x objectives (40x for normal hearts and 60x for ACTC1-variant heart and LMNA-R541C heart).

Techniques: Derivative Assay, Immunofluorescence, Isolation, Expressing

Journal: bioRxiv

Article Title: RNA polymerase loss by nuclear rupture drives LMNA cardiomyopathy

doi: 10.64898/2026.04.03.716433

Figure Lengend Snippet: A) Immunofluorescence for gamma-H2AX in cardiomyocytes expressing NLS-tdTomato and GFP-icGAS isolated from mice at day 14 post tamoxifen. Texts and icons below images indicate nuclear states. Scale bar: 5 μm. B) Nuclear gamma-H2AX intensity by nuclear states. Density and box plots (interquartile range): signal distribution of all affiliated nuclei. Circles: mean intensity within individual biological replicates (color coded). Statistics: P < 0.05 (*) or P≥0.05 (N.S.) from t-tests on linear regression-estimated means with mouse-clustered standard errors. Underlying data: 164 intact nuclei from 3 WT mice, 152 intact, 116 ruptured, 50 resealed nuclei from 3 Lmna CKO mice. C) Relationship between gamma-H2AX intensity and NLS-tdTomato intensity in nuclei of Lmna CKO cardiomyocytes (340 nuclei from 3 mice). Line: simple linear regression fit with 95% confidence interval. R: Pearson’s correlation coefficient. P: t-test p -value on linear regression-estimated means with mouse-clustered standard errors.

Article Snippet: Immunofluorescence images of human myocardial sections were acquired using a Yokogawa CSU-W1 Sora spinning disk confocal microscope with 40x or 60x objectives (40x for normal hearts and 60x for ACTC1-variant heart and LMNA-R541C heart).

Techniques: Immunofluorescence, Expressing, Isolation

Journal: bioRxiv

Article Title: RNA polymerase loss by nuclear rupture drives LMNA cardiomyopathy

doi: 10.64898/2026.04.03.716433

Figure Lengend Snippet: A) Model for the BANF1-ESCRT-III pathway driving nuclear membrane restoration on chromatin surface. B) Immunofluorescence for BANF1, LEMD2, CHMP7, CHMP4B, VPS4 and Lamin A/C in cardiomyocytes isolated at day 14 post tamoxifen. One binucleated cardiomyocyte per image, with nuclei zoomed in. Arrowheads: DNA protruded from rupture sites. Scale bar: 5 μm. C) Quantification of protruded chromatin colocalizing with BANF1-ESCRT-III pathway proteins. Graph shows mean percentage (bar) of biological replicates (points) with standard error ranges (error bars). Denominator: cardiomyocyte nuclei with protruded chromatin from 3 Lmna CKO mice. Denominator nucleus count: LEMD2 n=156, BANF1 n=170, CHMP4B n=158, VPS4 n=156, and CHMP7 n=174. D) Quantification of ruptured or resealed nuclei with LEMD2 or VPS4 localization on protruded chromatin. Denominator: ruptured or resealed nuclei from 3 Lmna CKO mice. Denominator nucleus count: LEMD2 n=171 nuclei (86 ruptured, 85 resealed), and VPS4 n=177 nuclei (119 ruptured, 58 resealed). Asterisk: p -value < 0.05 from Wald z-tests in binomial GLMMs estimating the probability of association. N=3 mice per genotype. E) Immunofluorescence for total VPS4, VPS4 EQ (Myc), and Lamin A/C in cardiomyocytes isolated at day 14 post tamoxifen. Scale bar: 5 μm. F) Immunofluorescence for total VPS4 in cardiomyocytes expressing GFP-icGAS and NLS-tdTomato at day 14 post tamoxifen. Arrowheads: icGAS puncta. Scale bar: 20 µm. G) Proportion of ruptured and resealed nuclei in Lmna CKO (n=4) or Lmna CKO ;VPS4 EQ (n=8) mice at day 14 post tamoxifen. Asterisk: p -value <0.05 from Wald z-tests in binomial GLMMs comparing the fraction of ruptured vs. resealed nuclei between genotypes. H) Gross morphology of hearts stained with Hematoxylin-Eosin at day 25 post tamoxifen. Scale bar: 1 mm. I) Fibrotic areas of myocardium positive for Masson’s Trichrome staining. Graph shows mean percentage (bar) of biological replicates (points) with standard error ranges (error bars). WT (n=5), WT; VPS4 EQ (n=5), Lmna CKO (n=5), Lmna CKO ;VPS4 EQ (n=4). Mean ± standard error. Asterisk: p -value <0.05 in one-way ANOVA with Tukey’s post-hoc test. J) Kaplan-Meier survival analysis. WT (n=16), WT; VPS4 EQ (n=23), Lmna CKO (n=19), Lmna CKO ;VPS4 EQ (n=25). Asterisk: p -value <0.05 in log-rank test. K) Summary of . Nuclear rupture is detrimental to hearts while rupture sealing is cardioprotective.

Article Snippet: Immunofluorescence images of human myocardial sections were acquired using a Yokogawa CSU-W1 Sora spinning disk confocal microscope with 40x or 60x objectives (40x for normal hearts and 60x for ACTC1-variant heart and LMNA-R541C heart).

Techniques: Membrane, Immunofluorescence, Isolation, Expressing, Staining

Journal: bioRxiv

Article Title: RNA polymerase loss by nuclear rupture drives LMNA cardiomyopathy

doi: 10.64898/2026.04.03.716433

Figure Lengend Snippet: A) Table lists specimens used in BANF1 immunofluorescence in . B) BANF1 intensity at the tip of individual nuclei in cardiomyocytes. Nuclear count: human normal heart n=200 from 4 specimen fields, human ACTC1 -mutant heart n=250 from 5 specimen fields, and LMNA p.R541C heart n=250 from 5 specimen fields.

Article Snippet: Immunofluorescence images of human myocardial sections were acquired using a Yokogawa CSU-W1 Sora spinning disk confocal microscope with 40x or 60x objectives (40x for normal hearts and 60x for ACTC1-variant heart and LMNA-R541C heart).

Techniques: Immunofluorescence, Mutagenesis

Journal: bioRxiv

Article Title: RNA polymerase loss by nuclear rupture drives LMNA cardiomyopathy

doi: 10.64898/2026.04.03.716433

Figure Lengend Snippet: A) Immunofluorescence for BANF1 and Desmin in human hearts. Top: heart biopsy from an individual with a LMNA p.R541C heterozygous mutation. Middle: autopsy heart from an individual with a pathogenic ACTC1 mutation. Bottom: apparently healthy autopsy heart. Inset: magnified areas enclosed by white squares. Scale bar: 20 μm. B) Three-dimensional reconstruction of BANF1 (green) and DNA (red) of nuclei (#1-4) in the LMNA p.R541C heart indicated in ( A ). Scale bar: 5 μm. C) Nuclei positive for nuclear-tip BANF1 per specimen field (normal heart, n=4 fields; ACTC1 heart, n=5 fields, LMNA -R541C heart, n=5 fields). Box: interquartile range. Asterisk: p -value <0.05 in Kruskal–Wallis test with Holm-corrected Wilcoxon tests. D) Model. Nuclear rupture drives LMNA -cardiomyopathy due to Pol II loss. Ruptured nuclei accumulate in mutant hearts due to frequent re-rupture of resealed nuclei.

Article Snippet: Immunofluorescence images of human myocardial sections were acquired using a Yokogawa CSU-W1 Sora spinning disk confocal microscope with 40x or 60x objectives (40x for normal hearts and 60x for ACTC1-variant heart and LMNA-R541C heart).

Techniques: Immunofluorescence, Mutagenesis

Journal: Discover Oncology

Article Title: Comprehensive pan-cancer analysis of FSCN1 as a marker for prognosis and immunity

doi: 10.1007/s12672-026-04949-7

Figure Lengend Snippet: Expression and subcellular localization of FSCN1. A FSCN1 expression in cancers in TCGA. B FSCN1 expression in cancers in TCGA + GTEx. C FSCN1 expression between tumor and paired normal samples in TCGA. D Immunofluorescence staining of the subcellular localization of FSCN1 in HPA database. (* p < 0.05; ** p < 0.01; *** p < 0.001, ns: no statistical differences)

Article Snippet: We used the immunofluorescence staining images of three human cancer cell lines (U251MG, A-431, and U2OS) to display the subcellular localization of FSCN1 in cells from the Human Protein Atlas (HPA) database ( https://www.proteinatlas.org/) [ ].

Techniques: Expressing, Immunofluorescence, Staining

Journal: medRxiv

Article Title: Acellular Adipose Tissue promotes anti-fibrotic remodeling in Phase II Study

doi: 10.64898/2026.02.04.26345214

Figure Lengend Snippet: a. Multiplex immunofluorescence images using the PhenoCycler of AAT Material profiling blood vessels (CD31 + ), adaptive immune cells: B cells (CD20 + ), CD3 + T Cells (CD4 + and CD8 + ), and antigen presenting cells (APCs) (HLA-DR + ) revealed tertiary lymphoid structures in AAT. Zoomed out image; scale bar=50 µm. Zoomed in image; scale bar=15 µm b. Multiplex immunofluorescence images reveal additional lymphoid aggregates in AAT material. Scale bar=15 µm c. Multiplex immunofluorescence images revealed Ki-67 + CD20 + proliferative B cells in aggregate with CD3 + cells. Scale bar=15 µm

Article Snippet: We performed cell phenotyping and specific cell type density quantification of Vectra opal multiplex immunofluorescence imaging using HALO® (RRID:SCR_018350) Image Analysis Platform Version v3.4 (Indica Labs, Inc.).

Techniques: Multiplex Assay, Immunofluorescence

Journal: medRxiv

Article Title: Acellular Adipose Tissue promotes anti-fibrotic remodeling in Phase II Study

doi: 10.64898/2026.02.04.26345214

Figure Lengend Snippet: a. Multiplex immunofluorescence images of AAT to investigate macrophage (CD68 + ) phenotype using classically-activated pro-inflammatory “M1” CD80 + and alternatively activated anti-inflammatory “M2” CD206 + markers. Zoomed out image; scale bar=100 µm. Zoomed in image; scale bar=20 µm b. CD68 + cell density in AAT material quantified by IF staining using HALO c. Macrophage polarization in AAT material quantified by IF staining using HALO. Populations of nonpolarized (CD68 + CD80 - CD206 - cells), classically-activated (CD68 + CD80 + CD206 - cells), alternatively activated (CD68 + CD206 + CD80 - cells), and double-polarized (CD68 + CD80 + CD206 + cells) represented as a percentage of CD68 + cells

Article Snippet: We performed cell phenotyping and specific cell type density quantification of Vectra opal multiplex immunofluorescence imaging using HALO® (RRID:SCR_018350) Image Analysis Platform Version v3.4 (Indica Labs, Inc.).

Techniques: Multiplex Assay, Immunofluorescence, Staining

Journal: medRxiv

Article Title: Acellular Adipose Tissue promotes anti-fibrotic remodeling in Phase II Study

doi: 10.64898/2026.02.04.26345214

Figure Lengend Snippet: a. Polarized light images of PSR staining of AAT material to investigate collagen network remodeling. Scale bar=10 µm b. Multiplex immunofluorescence staining of progenitor marker (CD34 + ), stromal marker (CD90 + ), and receptor for macrophage inhibitory factor (MIF) (CD74 + ) revealed pre-mature endothelial cells in AAT material important for angiogenesis. Scale bar=20 µm c. Multiplex immunofluorescence staining of progenitor marker (CD34 + ), stromal marker (CD90 + ), and receptor for macrophage inhibitory factor (MIF) (CD74 + ) to confirm presence of anti-fibrotic CD74 + ASCs (CD34 + CD90 + CD74 + ) in AAT Material. Scale bar=20 µm

Article Snippet: We performed cell phenotyping and specific cell type density quantification of Vectra opal multiplex immunofluorescence imaging using HALO® (RRID:SCR_018350) Image Analysis Platform Version v3.4 (Indica Labs, Inc.).

Techniques: Staining, Multiplex Assay, Immunofluorescence, Marker