primary human dermal lymphatic endothelial cells lecs  (PromoCell)

Journal: bioRxiv

Article Title: Oncogenic virus hijacks SOX18 pioneer function to enhance viral persistence

doi: 10.1101/2025.06.28.662102

Figure Lengend Snippet: A-C. HeLa cells expressing SOX18wt, mutants C240X (dominant-negative transactivation deficient) or HMGdel (DNA-binding deficient), or mCherry as a control, and thereafter infected with rKSHV.219 for 72h (KSHV-HeLa). A. IF images of the SOX18wt and mutants expressing cells labeled with anti-SOX18 antibody and a schematic of the constructs. B. Immunoblotting with anti-SOX18 antibody using β-actin as a loading control for normalization. C. RT-qPCR for the indicated viral genes in KSHV-HeLa. D. LECs infected with rKSHV.219 (KLECs) for 72 hours and treated with Sm4 or DMSO control for 24h and relative mRNA measured for indicated viral transcripts. Statistical significance was determined by one-way ANOVA with Dunnett correction for multiple comparisons; ns = non-significant. E-F. Uninfected LECs and KLECs 72h p.i. treated with DMSO or Sm4 for another 72h and E) labeled with anti-ARID1A and -BRG1 antibodies, nuclei were counterstained with Hoechst (33342), scale bar is 10µm, and F) immunoblotted for the indicated proteins and quantified as in B.

Article Snippet: Primary human dermal lymphatic endothelial cells LEC (Promocell; C-12216) were maintained in Microvascular MV-2 (Promocell; C-22121) medium supplemented with 5% fetal bovine serum, basic fibroblast growth factor, insulin-like growth factor 3, epidermal growth factor, gentamicin sulfate/amphotericin, ascorbic acid, and hydrocortisone; VEGF was not added.

Techniques: Expressing, Dominant Negative Mutation, Binding Assay, Control, Infection, Labeling, Construct, Western Blot, Quantitative RT-PCR

Journal: bioRxiv

Article Title: Oncogenic virus hijacks SOX18 pioneer function to enhance viral persistence

doi: 10.1101/2025.06.28.662102

Figure Lengend Snippet: A-C. A schematic of the inhibitor mode of action is shown in the top panels. CTG viability assay of uninfected LECs (LEC) or LECs infected with rKSHV.219 (KLEC) for 72h and treated with the indicated, increasing concentrations of BRG1 inhibitors A) ACBI1, B) FHT-1015 and C) PFI-3 (n=3), arrows indicate the selected concentration for following inhibitor assays. D. LECs and KLECs were treated with ACBI1, FHT-1015 and PFI-3 and treated with EdU for 4h before subjecting to EdU Click-It, imaged with Opera Phenix 20x and quantified from (n=6 independent replicates, and from each n=100 nuclei). Statistical significance was determined by one-way ANOVA with Dunnett correction for multiple comparisons, ns = non-significant. E-F. LECs infected with ΔORF50 and treated with Sm4, FHT-1015, or DMSO for 72h and processed for ATAC-seq. E. Peaks and heatmaps of the top 1000 genomic regions with reduced overall accessibility (dark blue maps) showing shared (dark blue line), unique to Sm4 (turquoise) and unique to FHT-1015 (purple) loss sites. F. Pearson’s analysis of the replicate (n = 3) samples. ns = non-significant.

Article Snippet: Primary human dermal lymphatic endothelial cells LEC (Promocell; C-12216) were maintained in Microvascular MV-2 (Promocell; C-22121) medium supplemented with 5% fetal bovine serum, basic fibroblast growth factor, insulin-like growth factor 3, epidermal growth factor, gentamicin sulfate/amphotericin, ascorbic acid, and hydrocortisone; VEGF was not added.

Techniques: Viability Assay, Infection, Concentration Assay