Journal: Oncology Letters

Article Title: Ferroptosis as a therapeutic vulnerability in MDM2 inhibition in dedifferentiated liposarcoma

doi: 10.3892/ol.2025.15015

Figure Lengend Snippet: Comparison of the expression levels of six ferroptosis-related genes among adipose, WDLPS and DDLPS tissues. Expression levels of (A) MDM2, (B) GPX4, (C) SLC7A11, (D) SLC3A2, (E) HMGCR and (F) DPP4. In each panel, data in the left graph are from GSE21050, GSE20559 and GSE41168 (U133 Plus 2.0) and data in the right graph are from GSE30929 and GSE35710 (U133A). *P<0.05, **P<0.01 and ***P<0.001 as indicated. Differences in levels among the different tissue types were analyzed using Kruskal-Wallis followed by Dunn's post hoc test. NS, not significant; WDLPS, well-differentiated liposarcoma; DDLPS, dedifferentiated liposarcoma; MDM2, mouse double minute 2 homolog; GPX4, glutathione peroxidase 4; SLC7A11, solute carrier family 7 member 11; SLC3A2, solute carrier family 3 member 2; HMGCR, 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase; DPP4, dipeptidyl peptidase-4.

Article Snippet: The antibodies used for immunoblotting were as follows: p53 (cat. no. 2527S; 1:1,000; Cell Signaling Technology, Inc.), MDM2 (cat. no. 86934S; 1:1,000; Cell Signaling Technology, Inc.), SLC7A11 (cat. no. A13685; 1:1,000; ABclonal Biotech Co., Ltd.), GPX4 (cat. no. A1933; 1:1,000; ABclonal Biotech Co., Ltd.), SLC3A2/CD98hc (cat. no. A3658; 1:1,000; ABclonal Biotech Co., Ltd.), 4EBP1 (cat. no. 9644; 1:1,000; Cell Signaling Technology, Inc.), phosphorylated (p)-4EBP1 (cat. no. 9459; 1:1,000; Cell Signaling Technology, Inc.), p70S6 (cat. no. 9202; 1:1,000; Cell Signaling Technology, Inc.), p-p70S6 (cat. no. 9205; 1:1,000; Cell Signaling Technology, Inc.) and β-actin (cat. no. ab6276; 1:1,000; Abcam).

Techniques: Comparison, Expressing

Journal: Oncology Letters

Article Title: Ferroptosis as a therapeutic vulnerability in MDM2 inhibition in dedifferentiated liposarcoma

doi: 10.3892/ol.2025.15015

Figure Lengend Snippet: Immunoblotting of dedifferentiated liposarcoma cell lines following treatment with either erastin or RSL3 alone or with nutlin-3 pre-treatment and cystine uptake assay of these cell lines following treatment with either single agent erastin or nutlin-3 or combination. (A) Representative western blots, (B) quantification of SLC3A2 expression and (C) cystine uptake in LPS853 cells. (D) Representative western blots, (E) quantification of SLC3A2 expression and (F) cystine uptake in NDDLS-1 cells. Results are presented as the mean ± standard error of the mean from three independent experiments. *P<0.05, **P<0.01 and ***P<0.001 vs. control (0 µM) or as indicated.

Article Snippet: The antibodies used for immunoblotting were as follows: p53 (cat. no. 2527S; 1:1,000; Cell Signaling Technology, Inc.), MDM2 (cat. no. 86934S; 1:1,000; Cell Signaling Technology, Inc.), SLC7A11 (cat. no. A13685; 1:1,000; ABclonal Biotech Co., Ltd.), GPX4 (cat. no. A1933; 1:1,000; ABclonal Biotech Co., Ltd.), SLC3A2/CD98hc (cat. no. A3658; 1:1,000; ABclonal Biotech Co., Ltd.), 4EBP1 (cat. no. 9644; 1:1,000; Cell Signaling Technology, Inc.), phosphorylated (p)-4EBP1 (cat. no. 9459; 1:1,000; Cell Signaling Technology, Inc.), p70S6 (cat. no. 9202; 1:1,000; Cell Signaling Technology, Inc.), p-p70S6 (cat. no. 9205; 1:1,000; Cell Signaling Technology, Inc.) and β-actin (cat. no. ab6276; 1:1,000; Abcam).

Techniques: Western Blot, Expressing, Control

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