Journal: bioRxiv

Article Title: Snapshot of in-cell protein contact sites reveals new host factors and hijacking of paraspeckles during influenza A virus infection

doi: 10.1101/2025.03.09.642134

Figure Lengend Snippet: (A) Cross-linking network of M2 and LAT1 (B) Log(RLU) normalised to siNT. Data are represented as mean ± SD (n = 3), with individual experiment means shown as circles. Statistical analysis was performed using ordinary one-way ANOVA. Statistical significance: *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. siSLC3A2 is replicated from . (C) Volcano plots showing fold-change (log2) versus significance (-log10 p-value) for SLC7A5 versus isotype control. Viral proteins are depicted in orange, SLC7A5 and SLC3A2 – in green. (D) Maximum intensity projection (MaxIP) of confocal images from A549 cells infected with WSN (Multiplicity of infection (MOI) 3) or mock at 12 hpi. Cells were stained for SLC7A5 (green), M2 (magenta), and nuclei (DAPI, cyan). Scale bars, 20 µm. (E) Pearson’s correlation coefficient quantification of SLC7A5 and M2 colocalisation in membrane regions. Analysis was performed in mock-infected cells (n = 64) and WSN-infected cells at 12 hpi (n = 119). Statistical significance was determined using the Mann-Whitney test (****p<0.0001). (F) Representative plane and intensity profile showing the co-localisation of SLC7A5 and M2 in A549 cells at 12 hpi.

Article Snippet: The following primary antibodies were used for western blot analysis: NONO mouse monoclonal (1:1,000, Proteintech, clone 2A2B10, catalogue no. 66361-1-Ig); NONO rabbit polyclonal (1:1,000, Proteintech, catalogue no. 11058-1-AP); SFPQ mouse monoclonal (1:1,000, Proteintech, clone 1G4A5, catalogue no. 67129-1-Ig); SFPQ rabbit polyclonal (1:1,000, Proteintech, catalogue no. 15585-1-AP); PSPC1 rabbit polyclonal (1:1,000, Proteintech, catalogue no. 16714-1-AP); SLC7A5 rabbit polyclonal (1:1,000, Proteintech, catalogue no. 28670-1-AP); SLC7A5 mouse monoclonal (1:1,000, Proteintech, clone 2G5H3, catalogue no. 67951-1-Ig); SLC3A2 rabbit polyclonal (1:1,000, Proteintech, catalogue no 15193-1-AP); SLC3A2 mouse monoclonal (1:1,000, Proteintech, clone 2B10F5, catalogue no. 66883-1-Ig); NP mouse monoclonal (1:1,000, Abcam, clone C43, catalogue no. ab128193); M2 mouse (1:1,000, ThermoFisher, clone 14C2, catalogue no. MA1-082); PB1, PB2, M1 (1: 1000, Abcam ab22396), NS1 (1:1,000, ThermoFisher, MA5-35909); Vinculin mouse (1:5000, Merck, catalogue no. V9131); Cyclophilin B rabbit monoclonal (1:5,000, CST, clone D1V5J, catalogue no. 43603).

Techniques: Control, Infection, Staining, Membrane, MANN-WHITNEY

Journal: Cell death & disease

Article Title: Targeting estrogen-regulated system x c - promotes ferroptosis and endocrine sensitivity of ER+ breast cancer.

doi: 10.1038/s41419-025-07354-0

Figure Lengend Snippet: Fig. 3 High level of SLC7A11/SLC3A2 is positively correlated with ERα and poor prognosis in breast cancer. A Analysis of the GEO transcriptome dataset GSE173300 showed an overlap of 2 genes between E2-regulated genes and ferroptosis-regulated genes in MCF-7 cells. B Analysis of the TCGA database showed a positive correlation between SLC7A11 and ESR1 mRNA levels in Luminal A and Luminal B breast cancer. C Western blot analysis showed SLC3A2 and SLC7A11 protein levels in ER+ breast cancer cell lines MCF-7, ZR-75-1, and T47D. D–G Analysis of the GEO databases showed that breast cancer patients with higher SLC7A11 (D, E) (GSE26304, GSE18229) and SLC3A2 (F, G) (GSE159956, GSE25055) expression levels had shorter overall survival (D, F) and relapse-free survival times (E, G). H, I, Immunohistochemistry (IHC) analysis showed that SLC7A11 (H) and SLC3A2 (I) protein levels were significantly higher in malignant breast tumors compared with benign breast tissues. Scale bars, 200 μm and 50 μm. SLC7A11 and SLC3A2 IHC staining were scored in breast tumor tissues from 31 patients. *P < 0.05, **P < 0.01.

Article Snippet: Rabbit anti-SLC3A2 polyclonal antibodies (cat#15193, Proteintech) were used at 1:300 dilution.

Techniques: Western Blot, Expressing, Immunohistochemistry

Journal: Cell death & disease

Article Title: Targeting estrogen-regulated system x c - promotes ferroptosis and endocrine sensitivity of ER+ breast cancer.

doi: 10.1038/s41419-025-07354-0

Figure Lengend Snippet: Fig. 5 SLC7A11 and SLC3A2 mediate ERα inhibition of ferroptosis in ER+ breast cancer cells. SLC7A11 (A) and SLC3A2 (B) were knocked down by two different shRNAs in MCF-7 and ZR-75-1 cell lines, respectively. Knockdown of SLC7A11 (C) or SLC3A2 (D) inhibited the growth of ER+ breast cancer cells. MCF-7 and ZR-75-1 cells with SLC7A11 or SLC3A2 knockdown, respectively, were subjected to colony formation assay. E Overexpression of SLC7A11 and SLC3A2 in MCF-7 and ZR-75-1 cells with ERα knockdown. F, G Overexpression of SLC7A11 and SLC3A2 decreased the increase of Lipid ROS level caused by ERα knockdown. Lipid ROS levels in indicated cells in Fig. 5E were detected by flow cytometry. H Overexpression of SLC7A11 and SLC3A2 rescued the cell growth inhibited by ERα antagonist Fulvestrant. MCF-7 and ZR-75-1 cells with SLC7A11/SLC3A2 overexpression were treated with 1 µM Fulvestrant for 72 h. I Ferroptosis inducer Erastin enhanced the sensitivity of ER+ breast cancer cells to Tamoxifen treatment. MCF-7 and ZR-75-1 cells subjected to colony formation assay were treated with 10 nM E2 in presence of with vehicle, 5 μM Tamoxifen, 10 μM Erastin or in combination for 72 h. Data are shown as Mean ± SEM (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001, ###P < 0.001; ns not significant.

Article Snippet: Rabbit anti-SLC3A2 polyclonal antibodies (cat#15193, Proteintech) were used at 1:300 dilution.

Techniques: Inhibition, Knockdown, Colony Assay, Over Expression, Cytometry

Journal: Cell death & disease

Article Title: Targeting estrogen-regulated system x c - promotes ferroptosis and endocrine sensitivity of ER+ breast cancer.

doi: 10.1038/s41419-025-07354-0

Figure Lengend Snippet: Fig. 6 SLC7A11 and SLC3A2 are upregulated in endocrine resistant breast cancer. A Analysis of the GEO database (GSE128458) showed the expression of many ferroptosis-related genes including SLC7A11 and SLC3A2 are significantly elevated in Tamoxifen-resistant MCF-7 cells compared to the parental MCF-7 cells. B Western blot analysis showed that SLC7A11 and SLC3A2 protein levels are significantly higher in Tamoxifen-resistant than in the parental MCF-7 cell line. C Kaplan–Meier analysis of the survival time of breast cancer treated with Tamoxifen in the TCGA database with high or low mRNA levels of both SLC7A11 and SLC3A2. D, E Immunohistochemistry analysis showed that SLC7A11 (D) and SLC3A2 (E) protein levels were significantly higher in endocrine therapy-resistant breast tumors compared with primary breast tumors. Scale bars, 200 μm, and 50 μm. SLC7A11 and SLC3A2 IHC staining were scored in breast tumor tissues from 25 patients. *P < 0.05, ***P < 0.001.

Article Snippet: Rabbit anti-SLC3A2 polyclonal antibodies (cat#15193, Proteintech) were used at 1:300 dilution.

Techniques: Expressing, Western Blot, Immunohistochemistry

Journal: Cell death & disease

Article Title: Targeting estrogen-regulated system x c - promotes ferroptosis and endocrine sensitivity of ER+ breast cancer.

doi: 10.1038/s41419-025-07354-0

Figure Lengend Snippet: Fig. 8 Working model. Estrogen inhibits ferroptosis via transcriptionally upregulation of the system xc −(SLC7A11 and SLC3A2) in ER+ breast cancer cells. Targeting SLC7A11 and SLC3A2 may offer a novel therapeutic option for patients with ER+ breast cancer, particularly those with endocrine resistance.

Article Snippet: Rabbit anti-SLC3A2 polyclonal antibodies (cat#15193, Proteintech) were used at 1:300 dilution.

Techniques: