Journal: Oxidative Medicine and Cellular Longevity

Article Title: Hypoxia Enhances Glioma Resistance to Sulfasalazine-Induced Ferroptosis by Upregulating SLC7A11 via PI3K/AKT/HIF-1 α Axis

doi: 10.1155/2022/7862430

Figure Lengend Snippet: Hypoxia promoted SLC7A11 expression via PI3K/AKT/HIF-1 α pathway. (a) Cell viability curves at different concentrations of SAS for 24 h. PX-478 can reverse the enhanced SAS resistance induced by hypoxia. (b) The IC 50 of SAS in hypoxia with PX-478 group was obviously low than that of hypoxia group. The bar graph showed mean ± SD of 3 independent experiments. (c) Protein expression level of HIF-1 α and SLC7A11 influenced by oxygen content and HIF-1 α inhibitor PX-478 measured by western blot. PX-478 markedly decreased HIF-1 α and SLC7A11 protein levels. (d) Protein expression level of PI3K, AKT, p-AKT, HIF-1 α , and SLC7A11 influenced by oxygen content and AKT inhibitor MK-2206 measured by western blot. AKT inhibitor MK-2206 along with hypoxia attenuated the expression level of p-AKT, HIF-1 α , and SLC7A11. (e) The relationship between HIF-1 α and SLC7A11 expression level in TCGA. SLC7A11 was positively correlated with HIF-1 α in 257 glioma tumor samples. ∗ P < 0.05 and ∗∗ P < 0.01.

Article Snippet: Reagent used contained SAS (HY-14655, MCE, USA), HIF-2 α -IN-4 (HY-136748, MCE, USA), PX-478 (HY-10231, MCE, USA), and MK-2206 (HY-10358, MCE, USA).

Techniques: Expressing, Western Blot

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Hypoxia Enhances Glioma Resistance to Sulfasalazine-Induced Ferroptosis by Upregulating SLC7A11 via PI3K/AKT/HIF-1 α Axis

doi: 10.1155/2022/7862430

Figure Lengend Snippet: PX-478 induced lipid peroxidation in the U87 and U251 cells. (a, b) Lipid peroxidation assessment in U87 and U251 cells after exposure to DMSO, PX-478, or PX-478+Lv-SLC7A11 measured by fluorescence microscope (a) and flow cytometry (b). (c) Quantitative analysis of the expression levels of MDA in the U87 and U251 cells after exposure to DMSO, PX-478, or PX-478+Lv-SLC7A11. MDA increased after treating with PX-478 and reversed in the presence of Lv-SLC7A11. (d) Transmission electron microscopy images of the U87 and U251 cells after exposure to DMSO, PX-478, or PX-478+Lv-SLC7A11. PX-478-treated cancer cells exhibited shrunken mitochondria with enhanced membrane density. Black arrow: normal mitochondria; white arrow: shrunken and high membrane density mitochondria. Quantitative analysis of mitochondrial length was listed on the right. (e, f) Cell apoptosis was detected by flow cytometry (e) and TUNEL fluorescence staining (f) in the U87 and U251 cells after exposure to DMSO, PX-478, SAS, PX-478+SAS, and PX-478+Lv-SLC7A11+SAS. The bar graph showed mean ± SD of 3 independent experiments. ∗ P < 0.05 and ∗∗ P < 0.01.

Article Snippet: Reagent used contained SAS (HY-14655, MCE, USA), HIF-2 α -IN-4 (HY-136748, MCE, USA), PX-478 (HY-10231, MCE, USA), and MK-2206 (HY-10358, MCE, USA).

Techniques: Fluorescence, Microscopy, Flow Cytometry, Expressing, Transmission Assay, Electron Microscopy, Membrane, TUNEL Assay, Staining

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Hypoxia Enhances Glioma Resistance to Sulfasalazine-Induced Ferroptosis by Upregulating SLC7A11 via PI3K/AKT/HIF-1 α Axis

doi: 10.1155/2022/7862430

Figure Lengend Snippet: PX-478 promoted the anticancer activity of SAS in vivo. (a) Xenografts derived from the U87 or U87/Lv-SLC7A11 cells which were treated with different drugs demonstrated tumor volume (b), tumor weight (c), body weight (d), and tumor tissue MDA (e). (f, g) Luciferase activities verified the decreased and increased tumor growth, respectively, in orthotopic glioma models established by the U87 or U87/Lv-SLC7A11 cells. (h) Survival curves showed the survival rates of the engrafted mice. (i) Immunohistochemical labelling showed that the downregulated expression of SLC7A11 after being treated with PX-478 and Lv-SLC7A11 could efficiently increase SLC7A11 expression in vivo. The bar graph showed mean ± SD of 3 independent experiments. ∗ P < 0.05 and ∗∗ P < 0.01.

Article Snippet: Reagent used contained SAS (HY-14655, MCE, USA), HIF-2 α -IN-4 (HY-136748, MCE, USA), PX-478 (HY-10231, MCE, USA), and MK-2206 (HY-10358, MCE, USA).

Techniques: Activity Assay, In Vivo, Derivative Assay, Luciferase, Immunohistochemical staining, Expressing

Journal: Cells

Article Title: Inhibition of Adipose Tissue Beiging by HIV Integrase Inhibitors, Dolutegravir and Bictegravir, Is Associated with Adipocyte Hypertrophy, Hypoxia, Elevated Fibrosis, and Insulin Resistance in Simian Adipose Tissue and Human Adipocytes

doi: 10.3390/cells11111841

Figure Lengend Snippet: INSTI treatment during beige adipocyte differentiation is associated with the inhibition of beige adipogenesis. ASCs were differentiated into beige adipocytes in the presence of DMSO (Ctrl), dolutegravir/DTG, raltegravir/RAL, or bictegravir/BIC or differentiated into white adipocytes with DMSO (Ctrl) for 14 days. ( A ) Relative mRNA expression levels of TMEM26, cluster of differentiation 137 (CD137), PRDM16, Cbp/P300-interacting-transactivator-1 (CITED1), and iodothyronine-deiodinase-2 (DIO2), were normalized against that of PPIA . The results are expressed as the mean ± SEM % of control cells ( n = 9, in duplicate). ( B ) Relative mRNA expression levels of uncoupling protein 1 (UCP1) was normalized against that of PPIA (left panel). The results are expressed as the mean ± SEM % of control cells ( n = 9, in duplicate). Whole-cell lysates were prepared and analyzed using immunoblots. Representative immunoblots of UCP1 and tubulin are shown (right panel). Densitometry analyses against tubulin as loading control were performed and expressed as a mean % of the value for control cells ± SEM ( n = 6). ( C ) Relative mRNA expression of fibroblast-growth factor-21 (FGF21) was normalized against that of PPIA (left panel). The results are expressed as the mean ± SEM % of control cells ( n = 9, in duplicate). After 14 days of differentiation with INSTIs, the levels of FGF21 in the culture medium (right panel) from the last 48 h were determined with ELISA ( n = 6, in duplicate). The results are expressed as the mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. control cells.

Article Snippet: In some experiments, ASCs were exposed simultaneously to INSTIs and to 5 μmol/L PX478 (Adooq Bioscience, Irvine, CA, USA).

Techniques: Inhibition, Expressing, Control, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Cells

Article Title: Inhibition of Adipose Tissue Beiging by HIV Integrase Inhibitors, Dolutegravir and Bictegravir, Is Associated with Adipocyte Hypertrophy, Hypoxia, Elevated Fibrosis, and Insulin Resistance in Simian Adipose Tissue and Human Adipocytes

doi: 10.3390/cells11111841

Figure Lengend Snippet: Treatment with INSTIs is associated with elevated extracellular matrix component expression in beige adipocytes. ASCs were differentiated into adipocytes, as described in . ( A ) Relative mRNA expression levels of collagens (COL1A2, COL6A1, COL1A1), lysyl oxidase (LOX), actin alpha 2, smooth muscle (ACTA2), TGFB1, and DDR1, were normalized against that of PPIA . The results are expressed as the mean ± SEM % of control cells ( n = 9, in duplicate). ( B ) Whole-cell lysates were prepared and analyzed using immunoblots. Representative immunoblots of COL6α1, COL1α1, and tubulin are shown. Densitometry analyses against tubulin as loading control were performed and expressed as a mean % of the value for control cells ± SEM ( n = 6). * p < 0.05, ** p < 0.01 vs. control cells.

Article Snippet: In some experiments, ASCs were exposed simultaneously to INSTIs and to 5 μmol/L PX478 (Adooq Bioscience, Irvine, CA, USA).

Techniques: Expressing, Control, Western Blot

Journal: Cells

Article Title: Inhibition of Adipose Tissue Beiging by HIV Integrase Inhibitors, Dolutegravir and Bictegravir, Is Associated with Adipocyte Hypertrophy, Hypoxia, Elevated Fibrosis, and Insulin Resistance in Simian Adipose Tissue and Human Adipocytes

doi: 10.3390/cells11111841

Figure Lengend Snippet: The inhibition of HIF1A by PX478 prevented the deleterious effects of INSTIs on beiging and fibrosis. ASCs were concomitantly treated with INSTIs and PX478 (HIF-1α inhibitor) throughout the differentiation process. The relative mRNA expression levels of ( A ) HIF1A , ( B ) UCP1, DIO2; PRDM16 , and ( C ) LOX were normalized against that of PPIA. The results are expressed as the mean ± SEM % of control cells ( n = 3, in duplicate). * p < 0.05, ** p < 0.01, *** p < 0.001 vs. control cells. # p < 0.05, ## p < 0.01 , ### p < 0.001 PX478-treated vs. non treated cells.

Article Snippet: In some experiments, ASCs were exposed simultaneously to INSTIs and to 5 μmol/L PX478 (Adooq Bioscience, Irvine, CA, USA).

Techniques: Inhibition, Expressing, Control

Journal: Nature Communications

Article Title: Treg-γδ T cell axis determines sexual dimorphism in hepatocarcinogenesis

doi: 10.1038/s41467-026-69603-w

Figure Lengend Snippet: Umap projection visualizing total ( a ) and gender-specific ( b ) of T cell clusters of human HCC scRNA-seq data (male, n = 16; female, n = 15). Umap plot showing the enrichment of representative cytotoxic ( c ) and exhausted genes ( d ) (male, n = 16; female, n = 15). e Single-cell transcription levels of representative genes in CD8 + T cells and γδ T cells between male and female (male, n = 16; female, n = 15). Single-cell transcription levels of S100A4 in T cell subsets ( f ) and Tregs ( g ). h Volcano plots of DEGs (male/female) in Tregs from scRNA-seq data. The red dots indicate upregulated genes. The blue dots indicate downregulated genes (male, n = 16; female, n = 15). i The relative mRNA expression of S100A4 in human HCC samples measured by qPCR (male, n = 20; female, n = 20). j The correlation analysis between the relative mRNA expression of S100A4 or FOXP3 in human HCC samples ( n = 37). k The overall survival of Foxp3 high expression ( n = 188) and Foxp3 low expression patients ( n = 194) from UCSC database. l Immunofluorescent staining of Foxp3, S100a4 and DAPI in orthotopic HCC. Scale bars, 50 μm (left). Quantification of Foxp3 + S100a4 + cells (right) ( n = 8). Mouse splenic Tregs were cultured in normoxia or hypoxia for 48 h. The relative mRNA expression of S100a4 by qPCR ( m ). The western blotting of Hif1α and S100a4 protein ( n ) ( n = 3). o Cut&Tag analysis was used to assess the interaction between Hif1α and S100a4 in Tregs. The relative expression of S100a4 with specific primers targeting the sequences located at +76 to +195 of S100a4 promoter locus by qPCR ( n = 4). Data represent three independent experiments. Data are shown as the mean ± SEM. P value of Pearson r correlation ( j ) or Log-rank (Mantel–Cox) test ( k ) or two-tailed unpaired Student’s t test ( i , l , m and o ). For additional details, see also Supplementary Fig. and Supplementary Data . Source data are provided as a Source Data file.

Article Snippet: For experiments with the Hif1α inhibitor PX-478 (Shanghai Yuanye Bio-Technology Co., Ltd), mice were treated by intraperitoneal injection at a dose of 30 mg/kg body weight twice a week after orthotopic implantation.

Techniques: Single Cell, Expressing, Staining, Cell Culture, Western Blot, Two Tailed Test