a549 cells  (ATCC)

Journal: Biochemistry and Biophysics Reports

Article Title: ATM inhibition restores IFN-γ sensitivity and induces ferroptosis in NSCLC via DNA damage response

doi: 10.1016/j.bbrep.2026.102568

Figure Lengend Snippet: IFN-γ responsiveness correlates DNA damage and repair responses in NSCLC cell lines (A) Relative cell viability of A549 or PC-9 treated with the indicated concentration of IFN-γ for 24 h are shown. Data are presented as mean ± SD. (B) Reactome analysis of the GSE180942 dataset was performed using differential CRISPR β-scores under IFN-γ treatment (Δβ = PC-9 − A549). Bars indicate the normalized enrichment score (NES) for each pathway indicated. Positive NES denotes pathways whose constituent genes are more essential in A549, whereas negative NES denotes pathways more essential in PC-9 upon IFN-γ treatment.

Article Snippet: The human non-small cell lung cancer cell lines PC-9 (kindly gifted from Dr. Kiura, Okayama University, Japan) and A549 (CCL-185, obtained from American Type Culture Collection) were used.

Techniques: Concentration Assay, CRISPR

Journal: Biochemistry and Biophysics Reports

Article Title: ATM inhibition restores IFN-γ sensitivity and induces ferroptosis in NSCLC via DNA damage response

doi: 10.1016/j.bbrep.2026.102568

Figure Lengend Snippet: Inhibition of ATM restores NSCLC cells to IFN-γ by inducing DNA damage response (A) Cell viability of A549 (left panel) or PC-9 (right panel) treated with IFN-γ (1000 ng/ml) and/or KU-55933 (10 μM) for 24 h are shown. Data are presented as mean ± SD. * p < 0.05. (B) Expression of γH2AX and b-Actin (loading control) in A549 (left panel) or PC-9 (right panel) cells treated with IFN-γ (1000 ng/ml) and/or KU-55933 (10 μM) for 24 h are shown.

Article Snippet: The human non-small cell lung cancer cell lines PC-9 (kindly gifted from Dr. Kiura, Okayama University, Japan) and A549 (CCL-185, obtained from American Type Culture Collection) were used.

Techniques: Inhibition, Expressing, Control

Journal: Biochemistry and Biophysics Reports

Article Title: ATM inhibition restores IFN-γ sensitivity and induces ferroptosis in NSCLC via DNA damage response

doi: 10.1016/j.bbrep.2026.102568

Figure Lengend Snippet: Inhibition of ATM in combination with IFN-γ induce ferroptosis in NSCLCs Cell viability of A549 (A) or PC-9 (B) treated with the indicated combination of IFN-γ (1000 ng/ml), KU-55933 (10 μM), Ferrostatin-1 (5 μM), and Liproxstatin-1 (5 μM) for 24 h are shown. Data are presented as mean ± SD. * p < 0.05.

Article Snippet: The human non-small cell lung cancer cell lines PC-9 (kindly gifted from Dr. Kiura, Okayama University, Japan) and A549 (CCL-185, obtained from American Type Culture Collection) were used.

Techniques: Inhibition

Journal: Cell Insight

Article Title: Viperin weakens IFN-I-induced immune activity by facilitating STAT1 degradation through E3 ligase UBE4A

doi: 10.1016/j.cellin.2026.100322

Figure Lengend Snippet: IFN-I promotes ubiquitination and degradation of UBR5 . (A) Western blot analysis of UBR5 protein levels in HT1080 (left) and A549 (right) cells treated with IFNα (1 000 IU/mL) for 3, 6, 9 and 12 h. (B, C) RT-qPCR analysis of Ubr5 and Ifit1 mRNA levels in HT1080 (B) and A549 (C) cells treated with IFNα (1 000 IU/mL) for 3, 6, 9 and 12 h. (D) Western blot analysis of Flag-UBR5 protein levels in HT1080 cells treated with IFNα for 6, 9 and 12 h (left) or dose of IFNα (right). (E) Immunoprecipitation-immunoblotting (IP-IB) analysis of the interaction between Viperin and UBE4A or UBR5 in HEK293T cells transfected with Myc-Viperin and then treated with IFNα (1 000 IU/mL) for 6 and 12 h. (F) Cycloheximide (CHX) chase assay of UBR5 in HT1080 cells treated with CHX (50 μg/mL) for 3 and 6 h. (G) Western blot analysis of Flag-UBR5 protein levels in HT1080 cells transfected Flag-UBR5 for 36 h, and then treated with or without MG132 (10 μM). (H-J) Western blot analysis of UBR5 protein levels in HT1080 cells treated with or without MG132 (10 μM) (H), MA (10 μM) (I) or PR619 (10 μM) (J). (K) IP analysis of ubiquitination of UBR5 in HT1080 cells treated with IFNα (1 000 IU/mL) for 6, 9 and 12 h. (L, M) IP analysis of K48-ubiquitination (48ub) (L) and K63-ubiquitination (63ub) (M) of UBR5 in HT1080 cells treated with or without IFNα (1 000 IU/mL). NS, not significant ( p > 0.05), one-way analysis of variance (ANOVA), two-tailed unpaired Student's t -test. Data are shown as means ± SD of four biological replicates (B, C), or are representative of three independent experiments (A, D-M).

Article Snippet: HEK293T, A549, HT1080, Vero, and RAW264.7 cells were obtained from the American Type Culture Collection (ATCC).

Techniques: Ubiquitin Proteomics, Western Blot, Quantitative RT-PCR, Immunoprecipitation, Transfection, Two Tailed Test

Journal: Cell Insight

Article Title: Viperin weakens IFN-I-induced immune activity by facilitating STAT1 degradation through E3 ligase UBE4A

doi: 10.1016/j.cellin.2026.100322

Figure Lengend Snippet: IFN-I promotes UBR5 degradation by up-regulating ITCH expression . (A) Potential interacting protein of UBR5 obtained from the PINA database. (B, C) RT-qPCR analysis of Itch mRNA levels in HT1080 (B), A549 and RAW264.7 (C) cells treated with IFN for 3, 6 and 9 h. (D) Western blot analysis of ITCH protein levels in HT1080 and A549 cells treated with IFNα (1 000 IU/mL) for 6, 9 and 12 h. (E, F) IP analysis of the interaction between endogenous UBR5 and ITCH in HT1080 cells (E) or primary mouse heart, liver, spleen, lung and kidney tissues (F). (G) Western blot analysis of UBR5 protein levels in Itch +/+ and Itch −/− cells. (H) Western blot analysis of UBR5 protein levels in HT1080 cells transfected with doses of Flag-ITCH. (I) RT-qPCR analysis of UBR5 mRNA levels in HT1080 cells transfected with doses of Flag-ITCH. (J) Western blot analysis of UBR5 protein levels in HT1080 cells transfected with or without Flag-ITCH, and then treated with MA (10 μM). (K) CHX chase assay of Flag-UBR5 in Itch +/+ and Itch −/− cells transfected with Flag-UBR5 for 36 h, and then treated with CHX (50 μg/mL) for 6 and 12 h. (L) CHX chase assay of UBR5 in HT1080 cells transfected with Flag-ITCH. (M) Western blot analysis of UBR5 protein levels in Itch +/+ and Itch −/− cells treated with IFNα for 6 and 12 h. (N) IP analysis of ubiquitination of UBR5 in Itch +/+ and Itch −/− cells. (O) IP analysis of ubiquitination of UBR5 in HT1080 cells transfected with doses of Flag-ITCH. (P) IP analysis of ubiquitination of UBR5 in Itch +/+ and Itch −/− cells treated with or without IFNα. (Q) RT-qPCR analysis of Ifit1 and Isg1 5 mRNA levels in cells as indicated, treated with IFNα for 6 and 12 h. NS, not significant ( p > 0.05), ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, one-way analysis of variance (ANOVA) (B, C, I), two-tailed unpaired Student's t -test (Q). Data are shown as means ± SD of four biological replicates (B, C, I, Q) or are representative of three independent experiments (D-H, J-P).

Article Snippet: HEK293T, A549, HT1080, Vero, and RAW264.7 cells were obtained from the American Type Culture Collection (ATCC).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Transfection, Ubiquitin Proteomics, Two Tailed Test

Journal: Cell Insight

Article Title: Viperin weakens IFN-I-induced immune activity by facilitating STAT1 degradation through E3 ligase UBE4A

doi: 10.1016/j.cellin.2026.100322

Figure Lengend Snippet: The multifunctional interfering peptide VS-IP1 enhances IFN-I-induced antiviral immune activity . (A) Western blot analysis of STAT1 protein levels in HT1080 cells treated with VS-IP1 and followed with IFNα for 9, 12 and 15 h. (B) RT-qPCR analysis of Ifit1 and Isg1 5 mRNA levels in HT1080 cells treated with VS-IP1 and IFNα. (C) RT-qPCR analysis of Ifit1 mRNA levels in A549 cells treated with IFNα and dose of VS-IP1. (D) Viral titers in supernatants from HT1080 cells treated with VS-IP1 and IFNα followed by VSV infection (MOI = 1.0), determined by TCID50 assay. (E) RT-qPCR analysis of VSV, H1N1, SeV and HSV viral RNA in HT1080 cells treated with VS-IP1 and IFNα, followed by viral infection. (F) RT-qPCR analysis of Ifit1 , Isg15 and Isg5 4 mRNA levels in Stat1 +/+ and Stat1 −/− HT1080 cells treated with VS-IP1 and IFNα. (G) Viral titers in supernatants from Stat1 +/+ and Stat1 −/− HT1080 cells treated with VS-IP1 followed by VSV infection (MOI = 1.0), determined by TCID50 assay. (H-K) RT-qPCR analysis of VSV (H), H1N1 (I), SeV (J) and HSV (K) viral RNA in Stat1 +/+ and Stat1 −/− cells treated with VS-IP1 and IFNα, followed by viral infection. (L) Viral titers in supernatants from Viperin +/+ and Viperin −/− HT1080 cells treated with VS-IP1 followed by VSV infection (MOI = 1.0), determined by TCID50 assay. (M) RT-qPCR analysis of VSV RNA in Viperin +/+ and Viperin −/− HT1080 cells treated with VS-IP1 and IFNα, followed by VSV infection. NS, not significant ( p > 0.05), ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, two-tailed unpaired Student's t -test (B-M). Data are shown as means ± SD of at least three biological replicates (B-M) or are representative of three independent experiments (A).

Article Snippet: HEK293T, A549, HT1080, Vero, and RAW264.7 cells were obtained from the American Type Culture Collection (ATCC).

Techniques: Activity Assay, Western Blot, Quantitative RT-PCR, Infection, TCID50 Assay, Two Tailed Test

Journal: Oncology Letters

Article Title: BUD23 is associated with malignancy and correlates with immune infiltration in NSCLC

doi: 10.3892/ol.2026.15608

Figure Lengend Snippet: BUD23 knockdown suppresses the proliferative and migration of NSCLC cells. (A) RT-qPCR was used to quantify BUD23 mRNA levels in HBE cells and a panel of NSCLC cell lines. Validation of BUD23 knockdown efficiency in (B) A549 and (C) H1299 cells by RT-qPCR. Assessment of cell viability in (D) A549 and (E) H1299 cells via CCK-8 assay and cell migration in (F) A549 and (G) H1299 cells by wound healing assay (magnification, ×10). CCK-8 assays of (H) A549 and (I) H1299 cells 24 h after BUD23 knockdown with or without subsequent co-culture with Jurkat T cells for 48 h in Transwell chambers. (J) Quantification of apoptosis in A549 and H1299 cells via Annexin V/PI flow cytometry. **P<0.01 and ***P<0.001 vs. HBE, NC, Con or as indicated. NSCLC, non-small cell lung cancer; RT-qPCR, reverse transcription-quantitative PCR; HBE, human bronchial epithelial; CCK-8, Cell Counting Kit-8; NC, negative control; Con, control; Si1/2, small interfering RNA targeting BUD23.

Article Snippet: The HBE [full name: HBE4-E6/E7 (Human Bronchial Epithelial Cells; cat. no. CRL-2078)] cell line, A549 lung adenocarcinoma cell line (cat. no. CCL-185), H1299 lung large cell carcinoma cell line (cat. no. CRL-5803), H460 lung large cell carcinoma cell line (cat. no. HTB-177) and Jurkat T cells (cat. no. TIB-152; a childhood T acute lymphoblastic leukemia T-cell line), were purchased from the American Type Culture Collection.

Techniques: Knockdown, Migration, Quantitative RT-PCR, Biomarker Discovery, CCK-8 Assay, Wound Healing Assay, Co-Culture Assay, Flow Cytometry, Reverse Transcription, Real-time Polymerase Chain Reaction, Cell Counting, Negative Control, Control, Small Interfering RNA

Journal: Oncology Letters

Article Title: BUD23 is associated with malignancy and correlates with immune infiltration in NSCLC

doi: 10.3892/ol.2026.15608

Figure Lengend Snippet: BUD23 knockdown significantly downregulates POLR2J expression in non-small cell lung cancer. (A) Venn diagram showing the intersection between LUAD GSEA core enrichment genes (key genes driving Hallmark DNA repair pathway enrichment) and LUSC GSEA core enrichment genes within the Hallmark DNA repair gene set, identifying 49 common genes. (B) Venn diagram showing the intersection between BUD23-correlated genes (correlation index >0.3) derived from the TCGA-LUAD cohort and BUD23-correlated genes (correlation index >0.3) derived from the TCGA-LUSC cohort via multi-gene correlation analysis, yielding 79 overlapping genes. (C) Venn diagram showing the secondary intersection between the 49 overlapping GSEA core enrichment genes (from panel A) and the 79 overlapping BUD23-correlated genes (from panel B), identifying 4 shared common genes: TAF6, POLR2J, RFC2 and VPS37D. (D) Validation of TAF6, POLR2J, RFC2 and VPS37D mRNA expression following BUD23 knockdown in A549 cells via reverse transcription-quantitative PCR. Cell-cycle analysis in (E) A549 and (F) H1299 cells following BUD23 knockdown. ***P<0.001 vs. Con. POLR2J, RNA polymerase II subunit J; LUAD, lung adenocarcinoma; GSEA, gene set enrichment analysis; LUSC, lung squamous cell carcinoma; TCGA, The Cancer Genome Atlas; TAF6, TATA-box binding protein associated factor 6; RFC2, replication factor C subunit 2; VPS37D, vacuolar protein sorting-associated protein 37D; Con, control; Si1, small interfering RNA targeting BUD23.

Article Snippet: The HBE [full name: HBE4-E6/E7 (Human Bronchial Epithelial Cells; cat. no. CRL-2078)] cell line, A549 lung adenocarcinoma cell line (cat. no. CCL-185), H1299 lung large cell carcinoma cell line (cat. no. CRL-5803), H460 lung large cell carcinoma cell line (cat. no. HTB-177) and Jurkat T cells (cat. no. TIB-152; a childhood T acute lymphoblastic leukemia T-cell line), were purchased from the American Type Culture Collection.

Techniques: Knockdown, Expressing, Derivative Assay, Biomarker Discovery, Reverse Transcription, Real-time Polymerase Chain Reaction, Cell Cycle Assay, Binding Assay, Control, Small Interfering RNA

Journal: iScience

Article Title: A comprehensive multi-omics and functional study of evolutionary adaptive responses to smoke

doi: 10.1016/j.isci.2026.115547

Figure Lengend Snippet: ALDH3A1 and NQO1 as guardians of epithelial survival and barrier function upon smoke exposure (A) A549 epithelial cell death upon 4 h of exposure to 0%–100% cigarette smoke extract in ALDH3A1 and NQO1 CRISPR-Cas9 knockout cells. Epithelial cell barrier function change in response to 0%–20% cigarette smoke extract exposure for 24 h, which was measured in real-time monitoring of electrical resistance and capacitance during and upon the establishment of epithelial monolayers using electric cell-substrate impedance sensing (ECIS) in ALDH3A1- (B) and NQO1-knockout (C) A549 cells. ∗ p value <0.05.

Article Snippet: For in vitro experiments human lung adenocarcinoma A549 cells were used (CCL-185, ATCC).

Techniques: CRISPR, Knock-Out, Electric Cell-substrate Impedance Sensing