k562 cell lines  (ATCC)

Journal: bioRxiv

Article Title: Ecological determinants of disease and immunity in myelodysplastic syndromes

doi: 10.64898/2026.05.05.720208

Figure Lengend Snippet: a, Bar plots of relative median fluorescence intensity (MFI) of granzyme B and perforin from intracellular cytokine staining of primary untreated MDS CD8 T cells after exposure to increasing doses of TGFβ. Relative MFI values are shown as mean ± SD. Data from n=6 MDS patients. Mann-Whitney U test used for statistical analysis (*p<0.05, **p<0.01). b, Top, volcano plot of differentially expressed genes in CD8 memory T cells close to (≤20µm) versus far from (>50µm) megakaryocytes. Bottom, schematic of CD8 memory T cells based on their distance to nearest megakaryocytes from spatial data. c, Relative percent spliced in (dPSI) value of differential 3’ splice site in mRNAs from SF3B1 mutant MDS patient bulk RNA-seq versus SF3B1 wild-type MDS. Highlighted names indicate mRNAs encoding proteins involved in TGFβ signaling. d, Diagram of proteins (in green) involved in TGFβ signaling which undergo aberrant RNA splicing in SF3B1 mutant MDS. e, Protein diagram of TGFBR1 with red indicating insertion of four amino acids (GPFS) encoded by the long mRNA isoform promoted in SF3B1 mutant cells. f, Alpha fold model of mutant TGFBR1 (green) overlaid with published crystal structure of wild-type TGFBR1 (gray). The GPFS amino acid insertion seen in SF3B1 mutant cells is indicated in red in the inset. g, Percentage (%) of phospho-SMAD2 Serine 465/467 (pS465/467) in K562 cells with the indicated genetic alterations in SF3B1 or TGFBR1. Mean ± SD. Two-way ANOVA. ***p<0.001, ****p<0.0001. h , Representative flow cytometry histograms of p-SMAD2 S465/467 from (g). i, Schematic of an SF3B1-mutant megakaryocyte with increased TGFβ production suppressing cytotoxic activity of nearby CD8 + T cells (cell-extrinsic effect); mutant cell simultaneously exhibits impaired TGFβ sensing (cell-intrinsic effect).

Article Snippet: The K562 human myeloid leukemia cell line was purchased from American Type Culture Collection (ATCC; #CCL-243).

Techniques: Fluorescence, Staining, MANN-WHITNEY, Mutagenesis, RNA Sequencing, Flow Cytometry, Activity Assay

Journal: bioRxiv

Article Title: Ecological determinants of disease and immunity in myelodysplastic syndromes

doi: 10.64898/2026.05.05.720208

Figure Lengend Snippet: (a) Sashimi plots of bulk RNA-sequencing data of an aberrant 3’ splice site usage in TGFBR1, MAP3K7 and SMURF2 mRNA in SF3B1 mutant acute myeloid leukemia (AML) (top; n=76 patients), SF3B1 wild-type (WT) AML (middle; n=739 patients), and normal bone marrow (bottom; n=26 patients). Red lines indicate SF3B1 mutant-specific junctions while black lines represent junction spanning reads in wild-type cells. The number of reads is listed, and the frequency of reads is in parentheses. b , Crystal structure of the short-isoform of TGFBR1 (grey) overlaid with that of the alpha-fold predicted model of SF3B1 mutant induced long-isoform (green). c, RT-PCR analysis of aberrant 3’ splice events in TGFBR1, MAP3K7, and SMURF2 in human isogenic K562 cells with knockin of SF3B1 K700E mutation. d , RT-PCR analysis of endogenous TGFBR1 , MAP3K7, and SMURF2 splicing in primary samples in healthy bone marrow control patients (n=5), patients with SF3B1 K700E mutant MDS (n=5) and non-splicing factor mutant patients with MDS (n=5). e , Sanger sequencing electropherogram of the top and bottom PCR products from gel-purified TGFBR1 RT-PCRs from MDS SF3B1 mutant patients shown in ( c ). The red box highlights the alternatively spliced sequence in the top band, which includes a 12-nucleotide insertion predominantly observed in SF3B1 K700E mutant MDS patients. The bottom band sequence is displayed below, showing the canonical exonic sequence. f , Western blot of K562 cells with SF3B1 mutation, TGFBR1 knockout (KO), or TGFBR1 KO with addback of TGFBR1 cDNA encoding the long or short isoform. g, Western blot of cytoplasmic, membrane, and soluble nuclear fractions of K562 cells with TGFBR1 KO alone or with overexpression of TGFBR1 cDNA encoding the long or short isoform.

Article Snippet: The K562 human myeloid leukemia cell line was purchased from American Type Culture Collection (ATCC; #CCL-243).

Techniques: RNA Sequencing, Mutagenesis, Reverse Transcription Polymerase Chain Reaction, Knock-In, Control, Sequencing, Purification, Western Blot, Knock-Out, Membrane, Over Expression

Journal: International Journal of General Medicine

Article Title: Lovastatin Targets LIPA to Induce ER Stress-Mediated Apoptosis in Acute Myeloid Leukemia: A Multi-Omics Study

doi: 10.2147/IJGM.S591023

Figure Lengend Snippet: The Effect of lovastatin on the activity, cell cycle, and apoptosis in AML cell lines K562 and THP-1 in vitro. ( A and B ) Effects of lovastatin and 4-PBA at different concentrations on cell activities of THP-1 and K562; ( C ) Effects of lovastatin and 4-PBA on cell cycles of THP-1 and K562; ( D ) Effects of lovastatin and 4-PBA on cell apoptosis of THP-1 and K562. AML, acute myeloid leukemia. Cells were treated with lovastatin (100 μM) or 4-PBA (5 μM) for 24 hours; **** p <0.0001.

Article Snippet: The human leukemia cell lines K562 (ATCC ® CCL-243TM) and THP-1 (ATCC ® TIB-202TM) were commercially obtained from Sangon Biotech (Shanghai, China).

Techniques: Activity Assay, In Vitro

Journal: International Journal of General Medicine

Article Title: Lovastatin Targets LIPA to Induce ER Stress-Mediated Apoptosis in Acute Myeloid Leukemia: A Multi-Omics Study

doi: 10.2147/IJGM.S591023

Figure Lengend Snippet: The mRNA expression levels of LIPA, ATF6, eIF2α, XBP1, IRE1A, DDIT3, HSP90AA1, and PERK. ( A-H ) The ER biomarkers expression in K562 cells ( I-P ). The ER biomarkers expression in THP-1 cells. Cells were treated with lovastatin (100 μM) or 4-PBA (5 μM) for 24 h; **** p <0.0001.

Article Snippet: The human leukemia cell lines K562 (ATCC ® CCL-243TM) and THP-1 (ATCC ® TIB-202TM) were commercially obtained from Sangon Biotech (Shanghai, China).

Techniques: Expressing

Journal: bioRxiv

Article Title: Feeder cell – the key component in producing scalable and fit NK cells for therapeutic use

doi: 10.64898/2026.04.21.718880

Figure Lengend Snippet: (A) Schematic representation of chimeric transgene constructs introduced into K562 feeder cells via lentiviral transduction. (B) Representative flow cytometry histograms showing surface expression of transgenes following lentiviral transduction of the K562 cell line. (C) Quantification of TGF-β1 from supernatants of irradiated feeder cells (illustrated) cultured alone or co-cultured with NK cells over a 72-hour period. Co-cultures were initiated at a density of 0.5 × 10 6 cells/mL, with NK cells added at an effector-to-target (E:T) ratio of 1:5. Supernatants were collected at the endpoint. Data are presented as mean ± standard deviation (SD) from n = 3 individual donors. Statistical analysis comparisons of feeder cells were made against K562 cells and another comparison between feeder cells alone vs co-cultures as indicated by lines between the pairs using two-sample paired t-test and two-sample unequal-variance t-test. p < 0.05 (*), p < 0.01 (**), p < 0.001 (***).

Article Snippet: The K562 chronic myelogenous leukemia cell line was purchased from ATCC (K-562; CCL-243) and used to generate the modified feeder cells.

Techniques: Construct, Transduction, Flow Cytometry, Expressing, Irradiation, Cell Culture, Standard Deviation, Comparison

Journal: bioRxiv

Article Title: Feeder cell – the key component in producing scalable and fit NK cells for therapeutic use

doi: 10.64898/2026.04.21.718880

Figure Lengend Snippet: NK cell cytotoxicity against K562 target cells was assessed in a 24-hour co-culture assay at varying effector-to-target (E:T) ratios. NK cells were collected for the assay at multiple timepoints: starting immediately after thawing (A), and at weeks 1 (B) and 4 (C), as well as during each sample’s the week precedin the last week of survival lon -term culture (D). (E) Responsiveness to IL-2 stimulation was evaluated in thawed NK cells using a 24-hour co-culture assay with K562 cells immediately after thawing. 500 IU/ml IL-2 was either added (+) or omitted (–) from the co-culture to assess its effect on cytotoxic activity. (F) Cytotoxicity against NALM-6 cells and (G) SH-Sy5y cells were measured one week after thawing using 24-hour co-culture assays. Each dot represents an individual donor; bars indicate mean values. Data are presented as mean ± SD from (A) n = 3, (B) n = 6, (C) n = 3, (D) n = 3, (E) n = 3, (F) n = 6 and (G) n = 6 individual donors. Data (E) points represent individual donors: donor A (circles), B (triangles) and C (squares). Statistical analysis comparisons were made against the Day 1, non-expanded NK cell condition and in (E) by comparing IL-2 added and omitted condition, using two-tailed paired t-test. p < 0.05 (*), p < 0.01 (**), p < 0.001 (***).

Article Snippet: The K562 chronic myelogenous leukemia cell line was purchased from ATCC (K-562; CCL-243) and used to generate the modified feeder cells.

Techniques: Co-culture Assay, Co-Culture Assay, Activity Assay, Two Tailed Test

Journal: bioRxiv

Article Title: Feeder cell – the key component in producing scalable and fit NK cells for therapeutic use

doi: 10.64898/2026.04.21.718880

Figure Lengend Snippet: (A) NK cells were co-cultured for 4 hours with (+) or without (–) K562 target cells and analyzed by flow cytometry after live/CD56+ gating. Surface expression of the activation marker CD69, degranulation marker CD107a, and apoptosis-inducing ligands FasL and TRAIL were measured on NK cells. Marker expression was quantified using median fluorescence intensity (MFI). (B) Secretion of inflammatory cytokines (IFN-γ and N -α) and cytotoxic granule components (granzyme B and perforin) were measured from co-culture supernatants using a multiplex bead-based assay. Data are presented as mean ± SD from (A) and (B) n = 3 individual donors. Data (A) and (B) points represent individual donors: donor A (circles), B (triangles) and C (squares). Statistical analysis comparisons were made against the Day 1, non-expanded NK cell condition unless otherwise indicated using two-tailed paired t-test. p < 0.05 (*), p < 0.01 (**), p < 0.001 (***).

Article Snippet: The K562 chronic myelogenous leukemia cell line was purchased from ATCC (K-562; CCL-243) and used to generate the modified feeder cells.

Techniques: Cell Culture, Flow Cytometry, Expressing, Activation Assay, Marker, Fluorescence, Co-Culture Assay, Multiplex Assay, Bead-based Assay, Two Tailed Test

Journal: Science Advances

Article Title: Healthy donor T cell receptors expand functional neoantigen recognition beyond patient vaccination

doi: 10.1126/sciadv.adz1156

Figure Lengend Snippet: ( A ) Proportion of neoantigens that were immunogenic out of the neoantigens pool tested on in vitro stimulated CD8 + from patient pre-vaccinated (and TILs) and post-vaccination PBMC. ( B ) Proportion of neoantigens that were immunogenic out of the neoantigens pool tested on in vitro stimulated CD8 + from healthy donors. ( C ) Individual IFN-γ ELISPOT wells showing responses of in vitro primed CD8 + T cells from healthy donors following stimulation with K562 aAPCs expressing the corresponding patient-specific HLA-A allele and pulsed with the indicated patient-specific neoantigen peptide (1 μg/ml). T cells were cocultured with aAPCs at a 10:1 effector-to-target ratio for 24 hours before ELISPOT development. Each panel represents a distinct well imaged independently from biological replicates under identical experimental conditions. ( D ) Representative tetramer plots of respective neoantigens from healthy donors and patients are shown. Dot plot representing the overall % tetramer positive of neoantigen-specific T cells for each immunogenic neoantigen from the healthy donors is depicted. ( E ) Clonotype tracking of T cell after TCR sequencing of naïve T cells; post-stimulated T cells; and tetramer-sorted T cells of PZP R>W , MOCOS S>L , and DDX19B M>V , respectively, from healthy donors.

Article Snippet: Human embryonic kidney (HEK) 293T (CRL-3216) and K562 (CCL-243) cell lines were purchased from the American Type Culture Collection.

Techniques: Immunopeptidomics, In Vitro, Enzyme-linked Immunospot, Expressing, Sequencing

Journal: Science Advances

Article Title: Healthy donor T cell receptors expand functional neoantigen recognition beyond patient vaccination

doi: 10.1126/sciadv.adz1156

Figure Lengend Snippet: ( A ) Fluorescence-activated cell sorting (FACS) analysis of DDX19B M>V tetramer staining of DDX19B M>V TCR-transduced T cells of patient A02 CD8 + T cells. DDX19B M>V TCR are derived from healthy donor (HD9) or patient A02. ( B ) Bar chart representing the CD137 + T cells of HD9 DDX19B M>V TCR-transduced T cells from A02 CD8 + against HLA-A*02:03 K562 aAPC. ( C ) Bar chart representing the CD137 + expression of HD9 DDX19B M>V TCR-transduced T cells cocultured with A02 tumor dissociates and third-party tumor dissociates (A01 tumor) with or without anti-MHC blocking. Dot plot of the live cell count ( D ) and % of tumor cell death measured by caspase-3/7 and PI staining ( E ) are shown for different T cells: tumor cells ratio for untransduced patient A02 T cells, HD9 DDX19B M>V TCR, and third-party TCR-transduced T cells. n.s., not significant.

Article Snippet: Human embryonic kidney (HEK) 293T (CRL-3216) and K562 (CCL-243) cell lines were purchased from the American Type Culture Collection.

Techniques: Fluorescence, FACS, Staining, Derivative Assay, Expressing, Blocking Assay, Cell Characterization