Journal: Nature Communications

Article Title: AAK1 activation-mediated iron trafficking drives ferroptotic cell death

doi: 10.1038/s41467-025-67523-9

Figure Lengend Snippet: A Protein levels associated with iron metabolism in various cancer cell lines treated with erastin for 12 h. A549, 8 μM; CAL51, 30 μM; HN6, 30 μM; HT1080, 4 μM; MDA-MB-231, 8 μM; MCF7, 30 μM. B , C Total ( B ) and divalent ( C ) iron levels of various cancer cell lines treated with erastin for 12 h. A549, 4 μM; CAL51, 20 μM; HN6, 20 μM; HT1080, 1 μM; MDA-MB-231, 2 μM; MCF7, 20 μM . D Schematic of the endocytosis assays. TFR1 in the cell surface were labelled with TFR1-specific antibodies at 4 °C followed by transferring cells to 37 °C to allow TFR1 internalization. The internalization levels of TFR1 will be established by antibody-labelled TFR1 from the cell surface evaluated by flow cytometry. This figure was created in BioRender. Lichao, L. (2025) https://BioRender.com/p1ntlvc . E Endocytosis assays of TFR1 in various cancer cell lines treated with erastin for 12 h. A549, 4 μM; CAL51, 20 μM; HN6, 20 μM; HT1080, 1 μM; MDA-MB-231, 2 μM; MCF7, 20 μM. F PKCβ was knocked out using single guide RNAs (sgRNAs) in MDA-MB-231 (left) and HT1080 (right) cells. G Total (left) and divalent (right) iron levels in the indicated MDA-MB-231 cells treated with erastin for 12 h. H , I Endocytosis assays of TFR1 in the indicated MDA-MB-231 ( H ) and HT1080 ( I ) cells treated with 2 μM or 1 μM erastin for 12 h, respectively. The cells with blue highlight were kept in 4 °C for dormancy. The cells with red highlight were transferred to 37 °C for internalization. The cells with grey highlight were labelled with non-specific antibody as isotype control. J , K Plasmids of PKCβⅠ or PKCβⅡ were transfected into PKCβ -knockout MDA-MB-231 ( J ) and HT1080 ( K ) cells, respectively. Endocytosis assays of TFR1 were performed in these cells treated with 2 μM or 1 μM erastin for 12 h, respectively. L Total (left) and divalent (right) iron levels in the indicated MDA-MB-231 cells treated with erastin for 12 h. A , F , J , K Data are representative of n = 3 biologically independent experiments. B , C , E , J , K Data are presented as means ± SD, n = 3 biologically independent experiments, unpaired two-tailed Student’s t test. G–I , L Data are presented as means ± SD, n = 3 biologically independent experiments, one-way ANOVA test.

Article Snippet: Human breast cancer cell MDA-MB-231 and MCF7, human fibrosarcoma cell HT1080, human Non-Small Cell Lung Cancer cell A549 were obtained from American Type Culture Collection.

Techniques: Transferring, Flow Cytometry, Control, Transfection, Knock-Out, Two Tailed Test

Journal: bioRxiv

Article Title: Broad-spectrum antiviral activity of the synthetic rocaglate zotatifin against multiple viruses

doi: 10.1101/2025.11.18.689102

Figure Lengend Snippet: HDFs ( A ) or A549 cells ( B ) were pretreated with zotatifin at doses of 15, 30, and 50 nM. Cells were then infected with MAYV AVR0565 strain and treated with the compound, as previously described. At 24 hours after infection, the levels of the E1 and nsP1 viral proteins were evaluated using Western blot analysis. One representative image of three independent assays is shown. Glyceroaldehyde-3-phosphate dehydrogenase (GAPDH) protein was used as a loading control. ( C ) A549 cells were treated or not with 50 nM zotatifin and then infected with MAYV. After 24 hours, MAYV E1 and nsP1 proteins were analyzed using an immunofluorescence assay. One representative microphotograph of at least 10 fields is shown. Scale bar: 50 μm. ( D ) A549 cells were treated with the indicated doses of zotatifin. After 48 hours of incubation, cell viability was evaluated using the MTT method. The data represent the mean ± standard deviation of one experiment with five replicates. Statistical analysis was carried out using a one-way ANOVA, followed by a Dunnett’s post hoc test. ns, not significant.

Article Snippet: Human dermal fibroblasts (HDFs, Cat. # PCS-201-012), human immortalized microglial cells (HMC3, Cat. # CRL-3304), human non-small cell lung cancer A549 cells (CCL-185), BSC40 (Cat. # CRL-2761), and Vero-E6 cells (CRL-1586) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Infection, Western Blot, Control, Immunofluorescence, Incubation, Standard Deviation

Journal: bioRxiv

Article Title: Broad-spectrum antiviral activity of the synthetic rocaglate zotatifin against multiple viruses

doi: 10.1101/2025.11.18.689102

Figure Lengend Snippet: A549 cells were treated or untreated with 50 nM zotatifin for 8 hours. Next, the relative transcription levels of IFNα ( A ), DDX58 ( B ), MxA ( C ), ISG15 ( D ), and TLR3 ( E ) genes were analyzed using RT-qPCR. Relative mRNA expression was calculated using the β-actin gene for normalization with the ΔΔCT method. Data represent the mean ± standard deviation of two independent experiments in triplicate. Data were analyzed using an unpaired t-test. ns, not significant; **, p < 0.01; ***, p < 0.001; and ****, p < 0.0001.

Article Snippet: Human dermal fibroblasts (HDFs, Cat. # PCS-201-012), human immortalized microglial cells (HMC3, Cat. # CRL-3304), human non-small cell lung cancer A549 cells (CCL-185), BSC40 (Cat. # CRL-2761), and Vero-E6 cells (CRL-1586) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Quantitative RT-PCR, Expressing, Standard Deviation

Journal: bioRxiv

Article Title: Broad-spectrum antiviral activity of the synthetic rocaglate zotatifin against multiple viruses

doi: 10.1101/2025.11.18.689102

Figure Lengend Snippet: A549 cells were pretreated with zotatifin, then infected with PR8-GFP ( A-D ), rVSV-GFP ( E-G ) or vaccinia ( H-I ) viruses at low (0.5) or high (5) multiplicity of infection (MOI). After 24 hours, the intensity of GFP, and the quantity of viral progeny in PR8-GFP and rVSV-GFP infected cells were assessed using flow cytometry, and a plaque-forming assay. Virus titers in the cell lysates of vaccinia virus infected cells were determined by plaque assay. The data represent the mean ± standard deviation of two independent experiments in triplicate. Statistical analysis was carried out using a one-way ANOVA followed by a Dunnett’s post hoc test or an unpaired t-test. ns, not significant; *, p < 0.05; **, p < 0.01; ***, p < 0.001; and ****, p < 0.0001.

Article Snippet: Human dermal fibroblasts (HDFs, Cat. # PCS-201-012), human immortalized microglial cells (HMC3, Cat. # CRL-3304), human non-small cell lung cancer A549 cells (CCL-185), BSC40 (Cat. # CRL-2761), and Vero-E6 cells (CRL-1586) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Infection, Flow Cytometry, Virus, Plaque Assay, Standard Deviation

Journal: bioRxiv

Article Title: Broad-spectrum antiviral activity of the synthetic rocaglate zotatifin against multiple viruses

doi: 10.1101/2025.11.18.689102

Figure Lengend Snippet: A549 cells were pretreated with zotatifin, then infected with PR8-GFP ( A-B ), rVSV-GFP ( C-D ), or vaccinia ( E-F ) viruses, as previously indicated. At the indicated hours after infection, levels of influenza A NP, VSV M, or vaccinia E3L proteins were evaluated using Western blot analysis. One representative image of three independent assays is shown. β-actin or GAPDH proteins were used as a loading controls.

Article Snippet: Human dermal fibroblasts (HDFs, Cat. # PCS-201-012), human immortalized microglial cells (HMC3, Cat. # CRL-3304), human non-small cell lung cancer A549 cells (CCL-185), BSC40 (Cat. # CRL-2761), and Vero-E6 cells (CRL-1586) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Infection, Western Blot

Journal: Scientific Reports

Article Title: Novel anti-LAG-3 antibody LBL-007 with anti-PD-1 blockade enhances antitumor immunity by promoting T cell-induced apoptosis

doi: 10.1038/s41598-025-21400-z

Figure Lengend Snippet: Viability of tumor cells co-cultured with activated Jurkat cells and incubated with different antibodies. ( A ) Viability of tumor cells co-cultured with Jurkat cells incubated with the indicated antibodies for 48 h. In the "A549 and Transwell" and "MGC-803 and Transwell" group, Jurkat cells were added to the upper compartment of the Transwell inserts and A549/MGC-803 were seeded in the lower compartment to avoid direct contact between cell types. The results are expressed as the mean ± SD from triplicates. * P < 0.05, ** P < 0.01. ( B ) Image of A549 cells and MGC-803 cells co-cultured with Jurkat cells after incubation with the indicated antibodies for 48 h by optical microscopy.

Article Snippet: Human non-small cell lung cancer cell lines A549 and PC-9, human esophagogastric adenocarcinoma cell lines HGC-27 and MGC-803, and the human T-lymphoblastic leukemia Jurkat cell line were obtained from the American Type Culture Collection (ATCC) (Manassas, VA, USA).

Techniques: Cell Culture, Incubation, Microscopy

Journal: Scientific Reports

Article Title: Novel anti-LAG-3 antibody LBL-007 with anti-PD-1 blockade enhances antitumor immunity by promoting T cell-induced apoptosis

doi: 10.1038/s41598-025-21400-z

Figure Lengend Snippet: LBL-007 and anti-PD-1 antibodies promote IL-2, IL-10, and TNF production. ( A ) ELISA of IL-2, IL-10, and TNF in the supernatants of the co-culture model after incubation with the indicated antibodies for 48 h. “A549” refers to the Jurkat-A549 co-culture model, "MGC-803" refers to the Jurkat-MGC-803 co-culture model. ( B ) qPCR of IL-2, IL-10, and TNF in cells from the Jurkat-A549 co-culture model (upper layer) or the Jurkat-MGC-803 co-culture model (lower layer) after incubation with the indicated antibodies for 48 h. The results are expressed as the relative expression of the gene normalized to the endogenous control and to the sample with the lowest expression set as 1 in each type of cell. The results are expressed as the mean ± SD from triplicates. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Human non-small cell lung cancer cell lines A549 and PC-9, human esophagogastric adenocarcinoma cell lines HGC-27 and MGC-803, and the human T-lymphoblastic leukemia Jurkat cell line were obtained from the American Type Culture Collection (ATCC) (Manassas, VA, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Co-Culture Assay, Incubation, Expressing, Control