normobaric hypoxia chambers  (BioSpherix)

Journal: International Journal of Oncology

Article Title: Hypoxia-induced exosomal CAMTA1 promotes radio-resistance in MDA-MB-231 cells by regulating NRG1 to mediate M2 macrophage polarization

doi: 10.3892/ijo.2026.5875

Figure Lengend Snippet: NRG1 expression is elevated in RAW264.7 macrophages co-cultured with 4T1 cells. (A) The expression of NRG1 was examined using next-generation RNA sequencing analysis. (B) The mRNA expression of NRG1 in RAW264.7 macrophages co-cultured with 4T1 cells was detected using reverse transcription-quantitative PCR. (C) The protein expression of NRG1 in RAW264.7 macrophages co-cultured with 4T1 cells was detected using immunoblotting analysis. *** P<0.001. NRG1, neuregulin 1.

Article Snippet: Blocked by 5% BSA (MilliporeSigma) for 1 h at room temperature, the membranes were incubated with primary antibodies against NRG1 (cat. no. 10527-1-AP; 1:1,000; Proteintech Group, Inc.), hypoxia-inducible factor-1α (HIF-1α; cat. no. 20960-1-AP; 1:2,000; Proteintech Group, Inc.), CD63 (cat. no. 25682-1-AP; 1:2,000; Proteintech Group, Inc.), CD81 (cat. no. 27855-1-AP; 1:1,000; Proteintech Group, Inc.), CD9 (cat. no. 20597-1-AP; 1:2,000; Proteintech Group, Inc.), ALIX (cat. no. 12422-1-AP; 1:5,000; Proteintech Group, Inc.) and GAPDH (cat. no. 10494-1-AP; 1:5,000; Proteintech Group, Inc.) overnight at 4°C.

Techniques: Expressing, Cell Culture, RNA Sequencing, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot

Journal: International Journal of Oncology

Article Title: Hypoxia-induced exosomal CAMTA1 promotes radio-resistance in MDA-MB-231 cells by regulating NRG1 to mediate M2 macrophage polarization

doi: 10.3892/ijo.2026.5875

Figure Lengend Snippet: Hypoxic MDA-MB-231 cells induces the M2 polarization of THP-1 macrophages. (A) The protein expression of HIF-1α was assessed using immunoblotting analysis. (B) The protein expression of HIF-1α in hypoxic MDA-MB-231/THP-1 co-cultures was assessed using immunoblotting analysis. (C) The cell migration was detected using Transwell. (D and E) The level of CD163 was detected using flow cytometry. The levels of (F) IL-10 and (G) IL-12 were detected using ELISA-related assay kits. ** P<0.01 and *** P<0.001. HIF-1α, hypoxia-inducible factor-1α.

Article Snippet: Blocked by 5% BSA (MilliporeSigma) for 1 h at room temperature, the membranes were incubated with primary antibodies against NRG1 (cat. no. 10527-1-AP; 1:1,000; Proteintech Group, Inc.), hypoxia-inducible factor-1α (HIF-1α; cat. no. 20960-1-AP; 1:2,000; Proteintech Group, Inc.), CD63 (cat. no. 25682-1-AP; 1:2,000; Proteintech Group, Inc.), CD81 (cat. no. 27855-1-AP; 1:1,000; Proteintech Group, Inc.), CD9 (cat. no. 20597-1-AP; 1:2,000; Proteintech Group, Inc.), ALIX (cat. no. 12422-1-AP; 1:5,000; Proteintech Group, Inc.) and GAPDH (cat. no. 10494-1-AP; 1:5,000; Proteintech Group, Inc.) overnight at 4°C.

Techniques: Expressing, Western Blot, Migration, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Oncology

Article Title: Hypoxia-induced exosomal CAMTA1 promotes radio-resistance in MDA-MB-231 cells by regulating NRG1 to mediate M2 macrophage polarization

doi: 10.3892/ijo.2026.5875

Figure Lengend Snippet: NRG1 expression is associated with M2 polarization and poor prognosis. (A) The expression of NRG1 in BC samples using TCGA database. (B) Spearman correlation analysis between NRG1 and CD163 expression in breast cancer samples. (C) The association of NRG1 expression with the overall survival in BC patients. NRG1, neuregulin 1; BC, breast cancer; TCGA, The Cancer Genome Atlas; GEPIA, Gene Expression Profiling Interactive Analysis.

Article Snippet: Blocked by 5% BSA (MilliporeSigma) for 1 h at room temperature, the membranes were incubated with primary antibodies against NRG1 (cat. no. 10527-1-AP; 1:1,000; Proteintech Group, Inc.), hypoxia-inducible factor-1α (HIF-1α; cat. no. 20960-1-AP; 1:2,000; Proteintech Group, Inc.), CD63 (cat. no. 25682-1-AP; 1:2,000; Proteintech Group, Inc.), CD81 (cat. no. 27855-1-AP; 1:1,000; Proteintech Group, Inc.), CD9 (cat. no. 20597-1-AP; 1:2,000; Proteintech Group, Inc.), ALIX (cat. no. 12422-1-AP; 1:5,000; Proteintech Group, Inc.) and GAPDH (cat. no. 10494-1-AP; 1:5,000; Proteintech Group, Inc.) overnight at 4°C.

Techniques: Expressing, Gene Expression

Journal: International Journal of Oncology

Article Title: Hypoxia-induced exosomal CAMTA1 promotes radio-resistance in MDA-MB-231 cells by regulating NRG1 to mediate M2 macrophage polarization

doi: 10.3892/ijo.2026.5875

Figure Lengend Snippet: Hypoxic MDA-MB-231 cells induces the M2 polarization of THP-1 macrophages via NRG1. (A) The mRNA expression of NRG1 was detected using RT-qPCR. (B) The transfection efficacy of Ov-NRG1 was detected using RT-qPCR and immunoblotting analysis. (C) The transfection efficacy of si-NRG1 was detected using RT-qPCR and immunoblotting analysis. (D) The level of IL-10 was detected using ELISA-related IL-10 assay kits. (E and F) The level of CD163 was detected using flow cytometry. (G) Following the treatment of anti-NRG1 blocking antibody, the level of IL-10 was detected using ELISA-related IL-10 assay kits. (H-I) Following the treatment of anti-NRG1 blocking antibody, the level of CD163 was detected using flow cytometry. * P<0.05, ** P<0.01 and *** P<0.001. NRG1, neuregulin 1; RT-qPCR, reverse transcription-quantitative PCR; Ov, overexpression; si, small interfering.

Article Snippet: Blocked by 5% BSA (MilliporeSigma) for 1 h at room temperature, the membranes were incubated with primary antibodies against NRG1 (cat. no. 10527-1-AP; 1:1,000; Proteintech Group, Inc.), hypoxia-inducible factor-1α (HIF-1α; cat. no. 20960-1-AP; 1:2,000; Proteintech Group, Inc.), CD63 (cat. no. 25682-1-AP; 1:2,000; Proteintech Group, Inc.), CD81 (cat. no. 27855-1-AP; 1:1,000; Proteintech Group, Inc.), CD9 (cat. no. 20597-1-AP; 1:2,000; Proteintech Group, Inc.), ALIX (cat. no. 12422-1-AP; 1:5,000; Proteintech Group, Inc.) and GAPDH (cat. no. 10494-1-AP; 1:5,000; Proteintech Group, Inc.) overnight at 4°C.

Techniques: Expressing, Quantitative RT-PCR, Transfection, Western Blot, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Blocking Assay, Reverse Transcription, Real-time Polymerase Chain Reaction, Over Expression

Journal: International Journal of Oncology

Article Title: Hypoxia-induced exosomal CAMTA1 promotes radio-resistance in MDA-MB-231 cells by regulating NRG1 to mediate M2 macrophage polarization

doi: 10.3892/ijo.2026.5875

Figure Lengend Snippet: Hypoxic MDA-MB-231 cells mediate the M2 polarization of THP-1 macrophages via exosomes. (A) The cell migration was detected using Transwell. Scale: 100 μ m. (B) The level of CD163 was detected using flow cytometry analysis. (C) The level for IL-10 was detected using ELISA-related IL-10 assay kits. (D) Transmission electron microscope was used for the inspection of exosomes. Scale: 100 μ m. (E) The analysis of exosome particle size. (F) The expressions of exosomal marker proteins were detected using immunoblotting analysis. (G) PKH67 staining was used to trace the exosomes. (H) The level of CD163 was detected using flow cytometry analysis. ** P<0.01 and *** P<0.001.

Article Snippet: Blocked by 5% BSA (MilliporeSigma) for 1 h at room temperature, the membranes were incubated with primary antibodies against NRG1 (cat. no. 10527-1-AP; 1:1,000; Proteintech Group, Inc.), hypoxia-inducible factor-1α (HIF-1α; cat. no. 20960-1-AP; 1:2,000; Proteintech Group, Inc.), CD63 (cat. no. 25682-1-AP; 1:2,000; Proteintech Group, Inc.), CD81 (cat. no. 27855-1-AP; 1:1,000; Proteintech Group, Inc.), CD9 (cat. no. 20597-1-AP; 1:2,000; Proteintech Group, Inc.), ALIX (cat. no. 12422-1-AP; 1:5,000; Proteintech Group, Inc.) and GAPDH (cat. no. 10494-1-AP; 1:5,000; Proteintech Group, Inc.) overnight at 4°C.

Techniques: Migration, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Transmission Assay, Microscopy, Marker, Western Blot, Staining

Journal: International Journal of Oncology

Article Title: Hypoxia-induced exosomal CAMTA1 promotes radio-resistance in MDA-MB-231 cells by regulating NRG1 to mediate M2 macrophage polarization

doi: 10.3892/ijo.2026.5875

Figure Lengend Snippet: Exosomal CAMTA1 promotes the M2 polarization of THP-1 macrophages. (A) The expression of CAMTA1 in BC samples was predicted using TCGA database. (B) The mRNA expression of CAMTA1 in MDA-MB-231 cells was detected using RT-qPCR. (C) The mRNA expression of CAMTA1 in exosomes was detected using RT-qPCR. (D) The mRNA expression of CAMTA1 in THP-1 macrophages was detected using RT-qPCR. (E) The transfection efficacy of sh-CAMTA1 was detected using RT-qPCR and immunoblotting analysis. (F) The mRNA expression of CAMTA1 in transfected MDA-MB-231 cells was detected using RT-qPCR. (G) The mRNA expression of CAMTA1 in transfected exosomes was detected using RT-qPCR. (H) The cell migration was detected using Transwell. (I and J) The level of CD163 was detected using flow cytometry analysis. (K) The level of IL-10 was detected using ELISA-related IL-10 assay kits. * P<0.05, ** P<0.01 and *** P<0.001. CAMTA1, Calmodulin-binding Transcription Activator 1; BC, breast cancer; TCGA, The Cancer Genome Atlas; RT-qPCR, reverse transcription-quantitative PCR; sh, short hairpin.

Article Snippet: Blocked by 5% BSA (MilliporeSigma) for 1 h at room temperature, the membranes were incubated with primary antibodies against NRG1 (cat. no. 10527-1-AP; 1:1,000; Proteintech Group, Inc.), hypoxia-inducible factor-1α (HIF-1α; cat. no. 20960-1-AP; 1:2,000; Proteintech Group, Inc.), CD63 (cat. no. 25682-1-AP; 1:2,000; Proteintech Group, Inc.), CD81 (cat. no. 27855-1-AP; 1:1,000; Proteintech Group, Inc.), CD9 (cat. no. 20597-1-AP; 1:2,000; Proteintech Group, Inc.), ALIX (cat. no. 12422-1-AP; 1:5,000; Proteintech Group, Inc.) and GAPDH (cat. no. 10494-1-AP; 1:5,000; Proteintech Group, Inc.) overnight at 4°C.

Techniques: Expressing, Quantitative RT-PCR, Transfection, Western Blot, Migration, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Binding Assay, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: International Journal of Oncology

Article Title: Hypoxia-induced exosomal CAMTA1 promotes radio-resistance in MDA-MB-231 cells by regulating NRG1 to mediate M2 macrophage polarization

doi: 10.3892/ijo.2026.5875

Figure Lengend Snippet: Exosomal CAMTA1 promotes tumor growth in vivo . (A) The transfection efficacy of Ov-CAMTA1 was detected using RT-qPCR and immunoblotting analysis. (B) The appearance of tumor. The tumor (C) volume and (D) weight. (E) The mRNA expression of CAMTA1 was detected using RT-qPCR. (F) The level of IL-10 was detected using ELISA-related IL-10 assay kits. (G) The level of CD163 was detected using immunohistochemistry analysis. (H) H&E staining. (I) The expression of Caspase 3 was detected using immunohistochemistry analysis. (J) The Spearman correlation analysis of CAMTA1 and NRG1. (K) The expression of NRG1 was detected using immunohistochemistry analysis. * P<0.05, ** P<0.01 and *** P<0.001. CAMTA1, Calmodulin-binding Transcription Activator 1; Ov, overexpression; RT-qPCR, reverse transcription-quantitative PCR; H&E, hematoxylin and eosin; NRG1, neuregulin 1.

Article Snippet: Blocked by 5% BSA (MilliporeSigma) for 1 h at room temperature, the membranes were incubated with primary antibodies against NRG1 (cat. no. 10527-1-AP; 1:1,000; Proteintech Group, Inc.), hypoxia-inducible factor-1α (HIF-1α; cat. no. 20960-1-AP; 1:2,000; Proteintech Group, Inc.), CD63 (cat. no. 25682-1-AP; 1:2,000; Proteintech Group, Inc.), CD81 (cat. no. 27855-1-AP; 1:1,000; Proteintech Group, Inc.), CD9 (cat. no. 20597-1-AP; 1:2,000; Proteintech Group, Inc.), ALIX (cat. no. 12422-1-AP; 1:5,000; Proteintech Group, Inc.) and GAPDH (cat. no. 10494-1-AP; 1:5,000; Proteintech Group, Inc.) overnight at 4°C.

Techniques: In Vivo, Transfection, Quantitative RT-PCR, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Staining, Binding Assay, Over Expression, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: bioRxiv

Article Title: Loss of KMT2D accelerates hypertrophic chondrocyte differentiation and senescence by increasing mitochondrial ROS production

doi: 10.64898/2026.03.18.712470

Figure Lengend Snippet: A) Immunofluorescence staining of the distal femur growth plate of E17.5 Kmt2d +/+ (top panel) and Kmt2d +/βGeo (bottom panel) embryos for hypoxia marker EF5 (green) and the hypertrophic marker collagen 10 (COLX; magenta). Nuclei were stained with DAPI (blue). Turquoise line indicates the length of the hypoxic region, yellow line the length of the hypertrophic region, and orange line the length of the overlap of the hypoxic and hypertrophic regions. Size of B) total growth plate shown in pixels, C) hypoxic (EF5+) region shown as the percentage of total growth plate, D) hypertrophic (COLX+) region shown as the percentage of total growth plate, and E) the overlap the hypoxic and hypertrophic region in the distal femur growth plate of E17.5 Kmt2d +/+ and Kmt2d +/ β Geo embryos defined as the proportion (in %) of the COLX+ region that is EF5+. Scale bar = 250 µm.

Article Snippet: Pregnant dams were weighed and the nitroimidazole hypoxia marker EF5 (Medchemexpress, HY-U00118) was administered through intraperitoneal injection with 10 mM EF5 at 1% of body weight (0.1 mL per 10 g weight) 17.5 days post-conception.

Techniques: Immunofluorescence, Staining, Marker

Journal: Frontiers in Immunology

Article Title: Integrating chronic inflammation and hypoxia: the potential role of HIF-1α in tumor behavior and therapy response in high-grade serous ovarian cancer

doi: 10.3389/fimmu.2026.1757708

Figure Lengend Snippet: (A) mRNA expression of HIF-1α in ovary (n=26) and omentum (n=21). (B) mRNA expression of HIF-1α in peritoneum at PS (n=23) and IDS (n=20). (C) Concentration of HIF-1α in ascites at PS (n=28), plasma at PS (n=28) and plasma at IDS (n=20). HGSOC, High-grade serous ovarian cancer; ACT, adjuvant chemotherapy; NACT, neoadjuvant chemotherapy; PS, primary surgery; IDS, interval debulking surgery; HIF-1α, Hypoxia-inducible factor 1-alpha.

Article Snippet: HIF-1α concentrations in ascites and plasma were quantified using a Human HIF-1α ELISA kit (CSB-E12112h, CUSABIO, Wuhan, China) following the manufacturer’s instructions.

Techniques: Expressing, Concentration Assay, Clinical Proteomics, Adjuvant

Journal: Frontiers in Immunology

Article Title: Integrating chronic inflammation and hypoxia: the potential role of HIF-1α in tumor behavior and therapy response in high-grade serous ovarian cancer

doi: 10.3389/fimmu.2026.1757708

Figure Lengend Snippet: (A) Concentration of HIF-1α in ascites at PS grouped by ESR (ESR ≤30 mm/h, n=5; ESR >30 mm/h, n=23). (B) Concentration of HIF-1α in plasma at PS grouped by ESR (ESR ≤30 mm/h, n=5; ESR >30 mm/h, n=23). (C) Concentration of HIF-1α in ascites at PS grouped by CRS (CRS 1, n=11; CRS 2, n=7; CRS 3, n=5). (D) Concentration of HIF-1α in plasma at PS grouped by CRS (CRS 1, n=11; CRS 2, n=7; CRS 3, n=5). PS, primary surgery; IDS, interval debulking surgery; ESR, erythrocyte sedimentation rate; CRS, chemotherapy response score; HIF-1α, Hypoxia-inducible factor 1-alpha.

Article Snippet: HIF-1α concentrations in ascites and plasma were quantified using a Human HIF-1α ELISA kit (CSB-E12112h, CUSABIO, Wuhan, China) following the manufacturer’s instructions.

Techniques: Concentration Assay, Clinical Proteomics, Sedimentation

Journal: Frontiers in Immunology

Article Title: Integrating chronic inflammation and hypoxia: the potential role of HIF-1α in tumor behavior and therapy response in high-grade serous ovarian cancer

doi: 10.3389/fimmu.2026.1757708

Figure Lengend Snippet: (A) mRNA expression of HIF-1α in peritoneum at PS stratified by PFS (PFS ≤30, n=17; PFS >30, n=6). (B) Kaplan-Meier survival curve of PFS according to peritoneal HIF-1α mRNA expression. HIF-1α low: peritoneal HIF-1α mRNA expression at PS is below optimal cutoff of 0.01124, n=17; HIF-1α high: peritoneal HIF-1α mRNA expression at PS is above optimal cutoff of 0.01124, n=6. (C) Kaplan-Meier survival curve of PFI according to peritoneal HIF-1α mRNA expression. HIF-1α low: peritoneal HIF-1α mRNA expression at PS is below median of 0.01124, n=9; HIF-1α high: peritoneal HIF-1α mRNA expression at PS is above median of 0.01124, n=11. PFS, progression free survival; PFI, platinum free interval; PS, primary surgery; HIF-1α, Hypoxia-inducible factor 1-alpha.

Article Snippet: HIF-1α concentrations in ascites and plasma were quantified using a Human HIF-1α ELISA kit (CSB-E12112h, CUSABIO, Wuhan, China) following the manufacturer’s instructions.

Techniques: Expressing