slc7a5 mouse monoclonal  (Proteintech)

Journal: iScience

Article Title: Dietary lipid overload creates a suppressive environment that impedes the antiviral functions of NK cells

doi: 10.1016/j.isci.2025.112396

Figure Lengend Snippet: Decreased function of NK cells in obese mice upon acute retrovirus infection For 13 weeks, C57BL/6 mice were fed with HFD or CD. At week 12, mice were infected i.v. with FV for 7 days or used as uninfected, naive control. Splenic NK cells (NK1.1 + CD3 − ) were analyzed as percentage (A) or absolute cell numbers (B). NK cells of naive mice are shown in gray, whereas NK cells from FV-infected mice are displayed in white. Statistically significant differences were analyzed by an unpaired t test. Data are presented as mean values ± SEM. FSC-A of NK cells from lean and obese mice are shown in (C). Heat maps show fold changes of percentages (D) and median fluorescence intensity (MFI) (E) of NK cells from naive or FV-infected mice fed with HFD relative to CD (HFD rel. to CD). Eight mice per group from two independent experiments were analyzed by an unpaired t test (KLRG1, CD98, IFNγ, GzmB %, MitoSpy, and puromycin) and Mann-Whitney test (GzmA and GzmB MFI). Statistically significant differences ( p ≤ 0.05) are indicated in the figure. See also Figure S3 .

Article Snippet: Rat monoclonal anti mouse CD98 (4F2) (clone RL388) , BioLegend , Cat# 128216; RRID: AB_2750549.

Techniques: Infection, Control, Fluorescence, MANN-WHITNEY

Journal: iScience

Article Title: Dietary lipid overload creates a suppressive environment that impedes the antiviral functions of NK cells

doi: 10.1016/j.isci.2025.112396

Figure Lengend Snippet: Tregs negatively regulate NK cells after short-term HFD upon acute retrovirus infection DEREG mice were fed with HFD for a total of 10 days. After 4 days, mice were infected with FV for seven days and depleted for Tregs by i.p. injections of DT (A). mRNA of splenocytes was isolated and analyzed for Il10 expression (B). Seven mice from two independent experiments were used for analyses. Spleen weight (C) and viral loads (D) were determined at 7 dpi (HFD 10 mice, HFD+DT 7 mice). NK cell numbers and percentages are displayed in (E). Proliferation was measured using KI67 (F). Expression of KLRG1 by NK cells is shown in (G). Percentages of degranulating CD107a + NK cells are shown in (H). NK cell cytotoxic capacity was determined by GzmB (I; MFI). Expression levels (MFI) of CD98 (J), MitoSpy (K), and Puromycin (L) in NK cells were analyzed from seven animals of two independent experiments. NK cells were isolated using magnetic beads and co-cultured with CFSE-stained YAC-1 tumor cells. In vitro killing was detected with BD Canto II (M). Five mice (HFD) and 7 mice (HFD+DT) from two independent experiments were used to analyze differences between groups. Statistically significant differences were analysed by an unpaired t test (B, C, F–I, K, and M) or Mann-Whitney test (D). IL-10 was neutralized by repetitive injections of anti-IL-10 monoclonal antibodies in short-term HFD mice (N). Splenic NK cells were isolated with magnetic beads and co-cultured with CFSE-stained YAC-1 tumor cells. In vitro killing was detected with BD Canto II. Nine mice per group from two independent experiments were used and analyzed by an unpaired t test. One outlier was removed (ROUT method). The significance threshold was set at 0.05. Data are presented as mean values ± SEM. DT: diphtheria toxin.

Article Snippet: Rat monoclonal anti mouse CD98 (4F2) (clone RL388) , BioLegend , Cat# 128216; RRID: AB_2750549.

Techniques: Infection, Isolation, Expressing, Magnetic Beads, Cell Culture, Staining, In Vitro, MANN-WHITNEY, Bioprocessing

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: 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: 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: Frontiers in Pharmacology

Article Title: Neuroprotective effects of ethanol extraction from Rubia yunnanensis Diels on chronic cerebral hypoperfusion: modulation of the System Xc-/GSH/GPX4 axis to alleviate oxidative stress and ferroptosis

doi: 10.3389/fphar.2025.1552228

Figure Lengend Snippet: Regulation of intracellular iron ion homeostasis and System Xc-/GSH/GHX4 pathway by RY-A. Expression of (A, B) TFRC, (A, C) FPN, (A, D) SLC7A11, (A, E) SLC3A2, (A, F) GSS, (A, G) GPX4 in rat hippocampal tissues was detected by Western blot. Data are expressed as mean ± SEM (n = 3). Compared to the Sham group, ****p < 0.0001; Compared to the 2VO group, ## p < 0.01, #### p < 0.0001. 2VO, 2-vessel occlusion; RY-A, Rubia yunnanensis alcohol extract.

Article Snippet: After that, the following antibodies were used: transferrin receptor antibody (1:5000; ab269513), anti-SLC7A11 antibody (1:1000; ab307601), anti-β-actin antibody (1:1000; ab8227), goat anti-rabbit IgG H&L (HRP) (1:10000; ab6721), and rabbit anti-mouse IgG H&L (HRP) (1:10000; ab6728), all from Abcam; anti-ferroportin/SLC40A1 antibody (1:500; 26601-1-AP) from Proteintech; and anti-CD98/SLC3A2 antibody (1:200; sc-136139), anti-GPX-4 antibody (1:100; sc-166120), and anti-GSS antibody (1:100; sc-365863) from Santa Cruz Biotechnology.

Techniques: Expressing, Western Blot

Journal: Frontiers in Pharmacology

Article Title: Neuroprotective effects of ethanol extraction from Rubia yunnanensis Diels on chronic cerebral hypoperfusion: modulation of the System Xc-/GSH/GPX4 axis to alleviate oxidative stress and ferroptosis

doi: 10.3389/fphar.2025.1552228

Figure Lengend Snippet: Regulation of intracellular iron ion homeostasis and System Xc-/GSH/GHX4 pathway by RY-A. Expression of (A, B) TFRC, (A, C) FPN, (A, D) SLC7A11, (A, E) SLC3A2, (A, F) GSS, (A, G) GPX4 in rat hippocampal tissues was detected by Western blot. Data are expressed as mean ± SEM (n = 3). Compared to the Sham group, ****p < 0.0001; Compared to the 2VO group, ## p < 0.01, #### p < 0.0001. 2VO, 2-vessel occlusion; RY-A, Rubia yunnanensis alcohol extract.

Article Snippet: After that, the following antibodies were used: transferrin receptor antibody (1:5000; ab269513), anti-SLC7A11 antibody (1:1000; ab307601), anti-β-actin antibody (1:1000; ab8227), goat anti-rabbit IgG H&L (HRP) (1:10000; ab6721), and rabbit anti-mouse IgG H&L (HRP) (1:10000; ab6728), all from Abcam; anti-ferroportin/SLC40A1 antibody (1:500; 26601-1-AP) from Proteintech; and anti-CD98/SLC3A2 antibody (1:200; sc-136139), anti-GPX-4 antibody (1:100; sc-166120), and anti-GSS antibody (1:100; sc-365863) from Santa Cruz Biotechnology.

Techniques: Expressing, Western Blot