peripheral blood mononuclear cells pbmcs  (Charles River Laboratories)

Journal: Frontiers in Genetics

Article Title: Single-cell transcriptomic profiling reveals CD27 + cytotoxic T Cell heterogeneity and exhaustion dynamics in colorectal cancer tumor microenvironment

doi: 10.3389/fgene.2026.1808171

Figure Lengend Snippet: Comparative induction of T cell exhaustion markers in colorectal cancer cell lines upon PBMC co-culture. Bar graph showing relative expression (fold change vs. monoculture baseline) of PDCD1, LAG3, HAVCR2, and CD27 in MSI-H cell lines (HCT116 and RKO) and MSS cell lines (HCT15 and SW480) following 48-h co-culture with activated PBMCs. Gray dashed line represents monoculture baseline (fold change = 1). MSI-H cell lines consistently demonstrated 2-3 times higher induction of all four exhaustion markers compared to MSS cell lines, with PDCD1 and LAG3 showing the greatest differential responses. Data represent mean values from three independent biological replicates analyzed by qRT-PCR with GAPDH normalization using the 2 -ΔΔCt method.

Article Snippet: Human peripheral blood mononuclear cells (PBMCs) were obtained commercially from the American Type Culture Collection (ATCC; catalog no. PCS-800-011) and used in accordance with the supplier’s guidelines.

Techniques: Co-Culture Assay, Expressing, Quantitative RT-PCR

Journal: iScience

Article Title: ALDH3A2-mediated fatty acid synthesis induces ferroptosis and AML drug resistance

doi: 10.1016/j.isci.2026.116202

Figure Lengend Snippet: Expression of ALDH3A2 in healthy people and patients with AML (A) The expression of ALDH3A2 in various tumors compared with normal tissues. Data are represented as mean ± SEM. (B) Expression of ALDH3A2 in 88 leukemia cell lines. (C) The survival curve of patients with high and low expression of ALDH3A2 (n [low expression] = 121, n [high expression] = 30). (D) The expression of protein ALDH3A2 in HL-60, HL-60/ADM, K562, K562/ADM, and bone marrow in patients. (E) The expression of ALDH3A2 RNA in bone marrow mononuclear cells of AML patients in the CR ( n = 9), C1NR ( n = 8), and R/R ( n = 10) group. Data are represented as mean ± SEM. (F) The expression of ALDH3A2 in bone marrow mononuclear cells of 3 AML patients at their initial and recurrent stage. (G) High/low expression of ALDH3A2 and molecular mutation distribution of 66 AML patients. Data are represented as mean ± SEM. Ns indicates no statistical difference between the two groups, ∗ indicates p < 0.05 between the two groups. Statistical significance was determined using unpaired two-tailed Student’s t tests for the comparison of two groups.

Article Snippet: ALDH3A2 expression in human leukemia cell lines and peripheral blood mononuclear cells , Human Protein Atlas website ( https://www.proteinatlas.org/ ) , .

Techniques: Expressing, Mutagenesis, Two Tailed Test, Comparison

Journal: iScience

Article Title: ALDH3A2-mediated fatty acid synthesis induces ferroptosis and AML drug resistance

doi: 10.1016/j.isci.2026.116202

Figure Lengend Snippet: ALDH3A2-V protects AML cells from ferroptosis and cytotoxicity induced by doxorubicin (A) ALDH3A2 mRNA relative expression in HL-60/ADM and K562/ADM cells after transfection of siRNA by electroporation. Data are represented as mean ± SEM. The specific data are shown in the supplemental table . (B) The lipid peroxidation degree of HL60ADM and K562ADM cells in the control (ctrl) and ALDH3A2 knockdown (KD) groups after the same dose of doxorubicin treatment for 48 h. (C) The content of ALDH3A2 protein in HL-60, K562, U937, and KG-1α in the ALDH3A2 overexpressing (OE) group and the control (CON) group. (D) The location of ALDH3A2-V overexpression. This representative image was selected from 3 independent replicate experiments, with 4 fields of view evaluated per replicate. (E) Lipid peroxidation in HL-60, K562, U937, and KG-1α in the ALDH3A2 OE group and the CON group under the same concentration of doxorubicin treatment. (F and G) Cell viability and its fitting curve of HL-60, K562, U937, and KG-1α in the ALDH3A2 OE group and the CON group under the gradient concentration of doxorubicin treatment. Data are represented as mean ± SEM. The specific data are shown in the supplemental table . Ns indicates no statistical difference between the two groups, ∗ indicates p < 0.05 between the two groups, and ∗∗ indicates p < 0.01 between the two groups. Statistical significance was determined using unpaired two-tailed Student’s t tests for the comparison of two groups.

Article Snippet: ALDH3A2 expression in human leukemia cell lines and peripheral blood mononuclear cells , Human Protein Atlas website ( https://www.proteinatlas.org/ ) , .

Techniques: Expressing, Transfection, Electroporation, Control, Knockdown, Over Expression, Concentration Assay, Two Tailed Test, Comparison

Journal: iScience

Article Title: ALDH3A2-mediated fatty acid synthesis induces ferroptosis and AML drug resistance

doi: 10.1016/j.isci.2026.116202

Figure Lengend Snippet: ALDH3A2 affects the sensitivity of AML cells to doxorubicin by altering 4-HNE and fatty acid content (A) The content of 4-HNE in cell lysate and culture supernatant of AML cells in the doxorubicin treatment (ADR) group and control (CON) group. Data are represented as mean ± SEM. The specific data are shown in the supplemental table . (B) The content of 4-HNE in HL-60, U937, and KG-1α of the overexpressing ALDH3A2 (OE) group and the CON group under the same concentration of doxorubicin treatment. Data are represented as mean ± SEM. The specific data are shown in the supplemental table . (C) OPLS-DA was performed on the measured fatty acids. Quantitative data for fatty acid and cell viability were collected from 3 independent experiments. (D) Variable Importance in Projection（VIP) value of different fatty acids; the difference is considered significant when the VIP>1. (E) Content of three different fatty acids in the CON and OE ALDH3A2 group. Data are represented as mean ± SEM. The specific data are shown in the supplemental table . (F) Cell viability of AML cells in heptadecanoic acid, oleic acid, and linoleic acid-pretreated groups and the CON group after the treatment of doxorubicin. Data are represented as mean ± SEM. The specific data are shown in the supplemental table . (G) Lipid peroxidation in AML cells in heptadecanoic acid-, oleic acid-, and linoleic acid-pretreated groups and the CON group after the treatment of doxorubicin. These images were selected from 3 independent replicates. Ns indicates no statistical difference between the two groups, ∗ indicates p < 0.05 between the two groups, ∗∗ indicates p < 0.01 between the two groups, and ∗∗∗ indicates p < 0.001 between the two groups. Statistical significance was determined using unpaired two-tailed Student’s t tests for the comparison of two groups.

Article Snippet: ALDH3A2 expression in human leukemia cell lines and peripheral blood mononuclear cells , Human Protein Atlas website ( https://www.proteinatlas.org/ ) , .

Techniques: Control, Concentration Assay, Two Tailed Test, Comparison

Journal: iScience

Article Title: ALDH3A2-mediated fatty acid synthesis induces ferroptosis and AML drug resistance

doi: 10.1016/j.isci.2026.116202

Figure Lengend Snippet: Effects of fatty acids and ALDH3A2 on plasma membrane fluidity and drug uptake (A) Laurdan staining showed different ratios of order and disorder phases in the control, heptadecanoic acid-, oleic acid-, or linoleic acid-treated groups. These representative images were selected from 3 independent replicate experiments, with 4 fields of view evaluated per replicate. (B) Flow cytometry was employed to quantify Cy5 fluorescence intensity in control, heptadecanoic acid-, oleic acid-, or linoleic acid-pretreated groups at 2- and 4-h intervals following doxorubicin exposure. The representative image was selected from 3 independent replicate experiments. Data are represented as mean ± SEM. The specific data are shown in the supplemental table . (C) Laurdan staining showed different ratios of order and disorder phases in the control and ALDH3A2 high groups. The mCherry fluorescent protein was co-expressed with ALDH3A2, which indicates successful transfection of the overexpression plasmid into the cells. These representative images were selected from 3 independent replicate experiments, with 4 fields of view evaluated per replicate. (D) U937 cells in the control group and ALDH3A2 high group were treated with Cy5-labeled doxorubicin. The mCherry fluorescent protein was co-expressed with ALDH3A2, which indicates successful transfection of the overexpression plasmid into the cells. These representative images were selected from 3 independent replicate experiments, with 4 fields of view evaluated per replicate. Ns indicates no statistical difference between the two groups, ∗ indicates p < 0.05 between the two groups, and ∗∗ indicates p < 0.01 between the two groups. Statistical significance was determined using unpaired two-tailed Student’s t tests for the comparison of two groups.

Article Snippet: ALDH3A2 expression in human leukemia cell lines and peripheral blood mononuclear cells , Human Protein Atlas website ( https://www.proteinatlas.org/ ) , .

Techniques: Clinical Proteomics, Membrane, Staining, Control, Flow Cytometry, Fluorescence, Transfection, Over Expression, Plasmid Preparation, Labeling, Two Tailed Test, Comparison

Journal: iScience

Article Title: ALDH3A2-mediated fatty acid synthesis induces ferroptosis and AML drug resistance

doi: 10.1016/j.isci.2026.116202

Figure Lengend Snippet: In in vivo experiments, AML with high expression of ALDH3A2 exhibits a poorer response to doxorubicin (A) Schematic diagram of the process for establishing the AML mouse model. (B) In vivo bioimaging shows the success of the model and the leukemia burden in mice before and after treatment. (C) Statistics of fluorescence values ( n = 5) on days 3 and 7 of the experiment. Data are represented as mean ± SEM. The specific data are shown in the supplemental table . (D) Survival curves of mice in different groups. (E) 4-HNE content in plasma of mice in different groups ( n = 5) on day 7. Data are represented as mean ± SEM. The specific data are shown in the supplemental table . (F) 4-HNE content in plasma of AML patients with low ( n = 5) and high ( n = 5) ALDH3A2 expression levels before and after receiving chemotherapy. Ns indicates no statistical difference between the two groups, ∗ indicates p < 0.05 between the two groups, and ∗∗ indicates p < 0.01 between the two groups. Statistical significance was determined using unpaired or paired two-tailed Student’s t tests for the comparison of two groups.

Article Snippet: ALDH3A2 expression in human leukemia cell lines and peripheral blood mononuclear cells , Human Protein Atlas website ( https://www.proteinatlas.org/ ) , .

Techniques: In Vivo, Expressing, Fluorescence, Clinical Proteomics, Two Tailed Test, Comparison

Journal: iScience

Article Title: ALDH3A2-mediated fatty acid synthesis induces ferroptosis and AML drug resistance

doi: 10.1016/j.isci.2026.116202

Figure Lengend Snippet: X24003 inhibits ALDH3A2 and enhances the cytotoxic effects of doxorubicin on resistant AML cells (A) OPLS-DA was performed on the measured fatty acid containment in the X24003 treatment group and control group. Quantitative data for fatty acid and cell viability were collected from 3 independent experiments. VIP value of different fatty acids; the difference is considered significant when the VIP>1. (B) X24003 exhibits a synergistic effect with doxorubicin in the elimination of doxorubicin-resistant AML cell lines HL-60ADM and K562ADM. (C) X24003 augments doxorubicin-induced lipid peroxidation in doxorubicin-resistant AML cell strains without affecting the parental cell lines. The representative image was selected from three independent replicate experiments. (D) In vivo bioimaging shows the success of the model and the leukemia burden in mice before and after treatment.

Article Snippet: ALDH3A2 expression in human leukemia cell lines and peripheral blood mononuclear cells , Human Protein Atlas website ( https://www.proteinatlas.org/ ) , .

Techniques: Control, In Vivo

Journal: iScience

Article Title: ALDH3A2-mediated fatty acid synthesis induces ferroptosis and AML drug resistance

doi: 10.1016/j.isci.2026.116202

Figure Lengend Snippet: HDAC2 binds to the promoter of the ALDH3A2 gene and regulates its expression (A) CUT&RUN indicates that HDAC2 binds to the promoter of the ALDH3A2 gene; the binding affinity is observed to decrease after treatment with chidamide. Data are represented as mean ± SEM. The specific data are shown in the supplemental table . (B) The expression of ALDH3A2 protein in AML cells following treatment with doxorubicin or chidamide. (C) Treatment of chidamide enhances lipid peroxidation induced by doxorubicin in AML cells. The representative image was selected from three independent replicate experiments. (D) Chidamide exhibits a synergistic effect with doxorubicin in the elimination of AML cells. Data are represented as mean ± SEM. The specific data are shown in the supplemental table . Ns indicates no statistical difference between the two groups, ∗ indicates p < 0.05 between the two groups, and ∗∗ indicates p < 0.01 between the two groups. Statistical significance was determined using unpaired two-tailed Student’s t tests for the comparison of two groups.

Article Snippet: ALDH3A2 expression in human leukemia cell lines and peripheral blood mononuclear cells , Human Protein Atlas website ( https://www.proteinatlas.org/ ) , .

Techniques: Expressing, Binding Assay, Two Tailed Test, Comparison

Journal: NAR Cancer

Article Title: Efficient gene disruption with CRISPR–Cas3 in human T cells

doi: 10.1093/narcan/zcag009

Figure Lengend Snippet: CRISPR–Cas3-engineered CAR-T cells retain cytotoxic function and exhibit reduced alloreactivity. ( A ) Flow cytometric analysis of TCRα/β and HLA–ABC expression in TRAC - or B2M -disrupted versus nondisrupted T cells after magnetic cell sorting. ( B ) CAR expression and residual TCRα/β or HLA–ABC expression in TRAC - or B2M -disrupted versus nondisrupted CAR-T cells assessed by flow cytometry. ( C ) Mixed lymphocyte reaction (MLR) assays evaluating alloreactivity of TRAC -disrupted CAR-T cells cocultured with irradiated allogeneic PBMCs, or B2M -disrupted CAR-T cells cocultured with allogeneic T cells. Data represent mean ± SD from triplicate wells ( n = 3). ( D ) Cytotoxic activity of TRAC - or B2M -disrupted CAR-T cells against GM2-positive or GM2-negative tumor cells (right panels), compared with non-genome-edited CAR-T cells processed under identical conditions (left panels). Data represent mean ± SD from triplicate wells ( n = 3). Statistical significance was determined using unpaired two-tailed Student’s t -test: * P < .05, ** P < .01, *** P < .001.

Article Snippet: Peripheral blood mononuclear cells (PBMCs) were obtained from Charles River Laboratories International, Inc.

Techniques: CRISPR, Expressing, FACS, Flow Cytometry, Irradiation, Activity Assay, Two Tailed Test