kat2a primary cell based screening assays cell culture ht1080 human fibrosarcoma cell line ht1080  (ATCC)

Journal: Bioactive Materials

Article Title: Multimodal profiling of CAR T cells against glioblastoma using a microengineered 3D tumor-on-a-chip model

doi: 10.1016/j.bioactmat.2026.01.003

Figure Lengend Snippet: Off-target cytotoxicity evaluation of CAR T cells using the 3D GOC system. A) Schematic representation of the differing cytolytic mechanisms of UTD, TV-13, and IL-13 CAR T cells against IL13Rα1 + HT-1080 tumor cells. Created with BioRender.com . B) Flow cytometric analysis confirming IL13Rα1 and mCherry (reporter gene) expression on IL13Rα1 + HT-1080 tumor cells. Antigen expression (IL13Rα1 or mCherry) on viable tumor cells shown in histograms: blue for IL13Rα1 + HT-1080 tumor cells and red for control tumor cells. The values within each histogram indicate the percentage of positive cells, with the mean fluorescence intensity (MFI) shown in parentheses. C) Microfluidic evaluation of off-target toxicities of T cells. (i) Representative tile images of tumor-stroma interface stained for actin cytoskeleton (green), showing differences in migration of IL13R1 + HT-1080 tumor cells (red) within the 3D GOC model across varying densities of UTD, TV-13 CAR, and IL-13 CAR T cells. (ii) Quantification of the migration distance of the IL13Rα1 + HT-1080 tumor cells in response to varying T cell concentrations. Data are represented as mean ± SD measured from three biological replicates ( n = 3) , T cell donors: DN18, DN28, and DN31, ∗ p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗ ∗p < 0.0001. Two-way ANOVA with Tukey's multiple comparisons test was utilized for statistical analysis. (iii) Bar graph showing the difference in nuclei per field of view (FOV) across different T cell densities, used as a measure of chain migration by IL13Rα1 + HT-1080 tumor cells. Data are represented as mean ± SD measured from three biological replicates ( n = 3) , T cell donors: DN18, DN28, and DN31, ∗ p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. Two-way ANOVA with Tukey's multiple comparisons test was utilized for statistical analysis, and (iv) Bar graph representing the percentage of T cells positive for intracellular cytokines in the presence of IL13Rα1 + HT-1080 tumor cells. Data are represented as mean ± SD measured from three biological replicates ( n = 3) , ∗ p < 0.05, ∗∗p < 0.01, ∗∗∗ ∗p < 0.0001. Two-way ANOVA with Tukey's multiple comparisons test was utilized for statistical analysis.

Article Snippet: HT-1080 Culture : Human fibrosarcoma cells (CCL-121, ATCC or HT-1080) were used to generate an off-target cell line (IL13Rα1 + HT-1080) expressing IL13Rα1-T2A-mCherry gene, which was single-sorted for the experiments described here.

Techniques: Gene Expression, Expressing, Control, Fluorescence, Staining, Migration

Journal: bioRxiv

Article Title: Nuclear blebs are composed of variable chromatin states but consistently enrich transcription initiation relative to elongation

doi: 10.64898/2026.03.10.710873

Figure Lengend Snippet: Example images of a nuclear bleb in HT1080 cells stably expressing NLS-GFP (green) and H2B-mCherry (gray). (A, B) Super plot nuclear bleb to body ratio (bleb/body) of NLS-GFP and H2B-mCherry (A) in cells in between nuclear ruptures, biological triplicates n = 22, and (B) in cells no nuclear rupture taken every hour, n = 5, 5, 3. Example images and super plots of nuclear bleb to body ratio for Hoechst 33342 DNA stain (cyan) and H3 (magenta) for cell lines and drug treatments (C) MEF WT n = 8, 15, 7, (D) MEF VPA n = 13, 11, 10, (E) HT1080 WT n = 10, (F) HT1080 VPA n = 8, 9, 13. White arrows denote the nuclear bleb. Mean ± s.e.m. is graphed. Statistical significance is denoted by *P<0.05, **P<0.01, ***P<0.001 or ns (not significant) via two-tailed paired Student’s t-test. Scale bars: 10 µm.

Article Snippet: Human fibrosarcoma HT1080 cells obtained from the American Tissue Culture Collection (ATCC) were cultured and passaged similarly.

Techniques: Stable Transfection, Expressing, Staining, Two Tailed Test

Journal: bioRxiv

Article Title: Nuclear blebs are composed of variable chromatin states but consistently enrich transcription initiation relative to elongation

doi: 10.64898/2026.03.10.710873

Figure Lengend Snippet: Examples of nuclear blebs in (A) MEF cells, (B) human HT1080 cells, and (C) human prostate cancer cell lines imaged via Hoechst (DNA) and euchromatin markers H3K27ac and H3K9ac. Super plots of nuclear bleb to body ratios biological triplicates in (A) MEF and (B) HT100 wild type (WT), increased euchromatin (VPA), decreased heterochromatin (DZNep), and lamin A knockdown (LA KD, MEF only). (C) Super plots of nuclear bleb to body ratios in human prostate cancer cell lines LNCaP n = 33, PC3 n = 30, and DU145 n = 29. Mean ± s.e.m. is graphed in A, B, and C. Statistical significance is denoted by * P <0.05, ** P <0.01, *** P <0.001 or ns (not significant) via One way Anova with post-hoc Turkey test. Scale bars: 10 µm.

Article Snippet: Human fibrosarcoma HT1080 cells obtained from the American Tissue Culture Collection (ATCC) were cultured and passaged similarly.

Techniques: Knockdown

Journal: bioRxiv

Article Title: Nuclear blebs are composed of variable chromatin states but consistently enrich transcription initiation relative to elongation

doi: 10.64898/2026.03.10.710873

Figure Lengend Snippet: Examples of nuclear blebs in (A) MEF cells, (B) human HT1080 cells, and (C) human prostate cancer cell lines imaged via Hoechst (DNA) and heterochromatin markers H3K27me3 and H3K9me2,3. Super plots of nuclear bleb to body ratios biological triplicates in (A) MEF and (B) HT100 wild type (WT), increased euchromatin (VPA), decreased heterochromatin (DZNep), and lamin A knockdown (LA KD, MEF only). (C) Super plots of nuclear bleb to body ratios in human prostate cancer cell lines LNCaP n = 10, 10, 9, PC3 n = 11, 11, 11, and DU145 n = 24. Mean ± s.e.m. is graphed in A, B, and C. Statistical significance is denoted by * P <0.05, ** P <0.01, *** P <0.001 or ns (not significant) via One way Anova with post-hoc Turkey test. Scale bars: 10 µm.

Article Snippet: Human fibrosarcoma HT1080 cells obtained from the American Tissue Culture Collection (ATCC) were cultured and passaged similarly.

Techniques: Knockdown

Journal: bioRxiv

Article Title: Nuclear blebs are composed of variable chromatin states but consistently enrich transcription initiation relative to elongation

doi: 10.64898/2026.03.10.710873

Figure Lengend Snippet: Example images and super plots of nuclear bleb to body ratio for DNA (cyan) and RNA Pol II pSer 2 (gray, elongation) and pSer5 (magenta, initiation) for cell lines (A) MEF n = 18, (B) HT1080 n = 10, (C) LNCaP n = 6, 10, 10, (D), PC3 n = 9, 7, 13, (E ) DU145 n = 12, 11, 11. (F) Graph summarizing all cell lines show a significant enrichment of RNA Pol II pSer5 relative to pSer2 in the nuclear bleb to body ratio. Mean ± s.e.m. is graphed. Statistical significance between pSer2 and pSer5 is denoted by * P <0.05, ** P <0.01, *** P <0.001 or ns (not significant) via two-tailed paired Student’s t -test. Scale bars: 10 µm.

Article Snippet: Human fibrosarcoma HT1080 cells obtained from the American Tissue Culture Collection (ATCC) were cultured and passaged similarly.

Techniques: Two Tailed Test