Journal: Annals of Hematology

Article Title: LncRNA PVT1 targets miR-143-3p to modulate endothelial cell function and thereby participate in deep vein thrombosis (DVT) of the lower limbs

doi: 10.1007/s00277-026-06833-4

Figure Lengend Snippet: Effects of si-PVT1 transfection on CoCl 2 –induced angiogenesis and inflammatory levels in HUVECs. A. Changes in vascular endothelial growth factor (VEGF) expression in HUVECs following CoCl 2 induction and si-PVT1 transfection. B. Effects of CoCl 2 induction and LncRNA PVT1 inhibition on fibroblast growth factor-2 (FGF-2) expression in HUVECs. Effects of different transfection treatments on inflammatory cytokine expression: TNF-α (C) and IL-6 (D) . * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: We used the following kits to assess the effects of different transfection treatments on endothelial cell angiogenesis and inflammatory levels: the Human VEGF ELISA Kit (97053ES, YENSEN, Shanghai), Human Fibroblast Growth Factor 2 (FGF2) ELISA Detection Kit (JL14546, JONLNBIO, Shanghai), Human TNF-α ELISA Kit (EK182, MULTI SCIENCES, Shanghai), and Human IL-6 ELISA Kit (EK106, MULTI SCIENCES, Shanghai).

Techniques: Transfection, Expressing, Inhibition

Journal: Annals of Hematology

Article Title: LncRNA PVT1 targets miR-143-3p to modulate endothelial cell function and thereby participate in deep vein thrombosis (DVT) of the lower limbs

doi: 10.1007/s00277-026-06833-4

Figure Lengend Snippet: Effects of miR-143-3p inhibitor on HUVEC function and inflammatory levels. Expression of LncRNA PVT1 (A) and miR-143-3p (B) in HUVECs following miR inhibitor transfection. C. Effect of miR inhibitor on viability of damaged HUVECs. D. Apoptotic changes in damaged HUVECs after miR inhibitor transfection. E. Effect of miR inhibitor on apoptosis-related gene expression in damaged HUVECs. F. Changes in VEGF and FG-2 expression in damaged HUVECs following miR inhibitor transfection. G. Effect of miR inhibitor on expression of inflammatory cytokines TNF-α and IL-6 in damaged HUVECs. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: We used the following kits to assess the effects of different transfection treatments on endothelial cell angiogenesis and inflammatory levels: the Human VEGF ELISA Kit (97053ES, YENSEN, Shanghai), Human Fibroblast Growth Factor 2 (FGF2) ELISA Detection Kit (JL14546, JONLNBIO, Shanghai), Human TNF-α ELISA Kit (EK182, MULTI SCIENCES, Shanghai), and Human IL-6 ELISA Kit (EK106, MULTI SCIENCES, Shanghai).

Techniques: Expressing, Transfection, Gene Expression

Journal: Cell Death Discovery

Article Title: Mitochondrial retrograde signaling initiates HIF-1α/BNIP3/NIX-mediated mitophagy in Tibetan high-altitude adaptation

doi: 10.1038/s41420-025-02933-8

Figure Lengend Snippet: A , B Western blot analysis of HIF-1α, BNIP3, NIX, MFN1, DRP1, LC3B, and p62 protein levels in M7/8+shCTR ( n = 4), M9a+shCTR ( n = 4), and M9a+sh EPAS1 ( n = 4) cells under hypoxia. Grayscale analysis of target protein levels was normalized to β-Tubulin. C Autophagosomes were analyzed in M7/8+shCTR ( n = 2), M9a+shCTR ( n = 2), and M9a+sh EPAS1 ( n = 2) cells using TEM under hypoxia. Scale bar: 1 μm. D Autophagic flux was assessed by quantifying the colocalization of LC3B (green) and Mitotracker (red) (marking mitophagosomes) using immunofluorescence in M7/8+shCTR ( n = 4), M9a+shCTR ( n = 4), and M9a+sh EPAS1 ( n = 4) cells under hypoxia. DAPI (blue) stains the nucleus. Scale bar: 20 μm. E Cytoplasmic free mtDNA was determined by staining the double-stranded DNA (dsDNA) (green) and Mitotracker (red) using immunofluorescence in M7/8+shCTR ( n = 4), M9a+shCTR ( n = 4), and M9a+sh EPAS1 ( n = 4) cells under hypoxia. DAPI (blue) stains the nucleus. dsDNA that does not colocalize with either the nucleus or mitochondria is considered cytoplasmic free mtDNA. Scale bar: 20 μm. F Western blot analysis of NLRP3, IL1β, cGAS, STING, P65, and P-P65 protein levels in M7/8+shCTR ( n = 4), M9a+shCTR ( n = 4), and M9a+sh EPAS1 ( n = 4) cells under hypoxia. Grayscale analysis of target protein levels was normalized to β-Tubulin. G Inflammatory cytokines IL6, IL1β, and TNF-α in the culture medium were measured in M7/8+shCTR ( n = 4), M9a+shCTR ( n = 4), and M9a+sh EPAS1 ( n = 4) cells under normoxic and hypoxic conditions using an ELISA kit. All cells were cultured under normoxia (21% O₂) or hypoxia (1% O₂) for 48 h.#1–4 represent individual samples. Data are presented as the mean ± SD of three independent experiments. One-way ANOVA followed by Tukey’s test; * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Human IL-6, IL-1β, and TNF-α levels were measured using ELISA kits (LiankeBio, China) according to the manufacturer’s instructions.

Techniques: Western Blot, Immunofluorescence, Staining, Enzyme-linked Immunosorbent Assay, Cell Culture

Journal: Cell Death Discovery

Article Title: Mitochondrial retrograde signaling initiates HIF-1α/BNIP3/NIX-mediated mitophagy in Tibetan high-altitude adaptation

doi: 10.1038/s41420-025-02933-8

Figure Lengend Snippet: A Total ROS and mtROS contents were determined in the M9a+shCTR ( n = 4), M9a+sh EPAS1 ( n = 4) and M9a+sh EPAS1 -NAC ( n = 4). The mtROS and total ROS contents in the M9a+sh EPAS1 and M9a+sh EPAS1 -NAC cells were normalized to those in the M9a+shCTR cells. B Western blot analysis of HIF-1α, BNIP3, NIX, LC3B, and p62 protein levels in the three cell lines under hypoxia. Grayscale analysis of target protein levels was normalized to β-Tubulin. C Autophagic flux was quantified by colocalizing LC3B (green) and Mitotracker (red) (mitophagosomes) using immunofluorescence in the three cell lines under hypoxia. DAPI (blue) stains the nucleus. Scale bar: 20 μm. D – F Mitochondrial OCR (basal, ATP-linked, maximal), MMP, and ATP content were measured in the three cell lines under hypoxia. MMP and ATP levels in M9a+sh EPAS1 and M9a+sh EPAS1 -NAC cells were normalized to those in M9a+shCTR cells. G CCK-8 assay showing cell viability in the three cell lines under hypoxia. H Cell apoptosis was determined using Annexin V-FITC staining in the three cell lines under hypoxia. I Western blot analysis of C-Caspase-3 and BAX protein levels in the three cell lines under hypoxia. Grayscale analysis of target protein levels was normalized to β-Tubulin. J Cytoplasmic free mtDNA was determined by staining dsDNA (green) and Mitotracker (red) using immunofluorescence in the three cell lines under hypoxia. DAPI (blue) stains the nucleus. dsDNA that does not colocalize with either the nucleus or mitochondria is considered cytoplasmic free mtDNA. Scale bar: 20 μm. K Western blot analysis of NLRP3, IL1β, cGAS, and STING protein levels in the three cell lines under hypoxia. Grayscale analysis of target protein levels was normalized to β-Tubulin. L Inflammatory cytokines IL6, IL1β, and TNF-α in the culture medium supernatant were measured in the three cell lines under hypoxia using an ELISA kit. The three cell lines, M9a+shCTR ( n = 4), M9a+sh EPAS1 ( n = 4), and M9a+sh EPAS1 -NAC ( n = 4), were treated with hypoxia (1% O₂) for 48 h, with NAC treatment (10 mM) administered during the last 24 h.#1–4 represent individual samples. Data are presented as the mean ± SD of three independent experiments. One-way ANOVA followed by Dunnett’s test; * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Human IL-6, IL-1β, and TNF-α levels were measured using ELISA kits (LiankeBio, China) according to the manufacturer’s instructions.

Techniques: Western Blot, Immunofluorescence, CCK-8 Assay, Staining, Enzyme-linked Immunosorbent Assay

Journal: Cell Death Discovery

Article Title: Mitochondrial retrograde signaling initiates HIF-1α/BNIP3/NIX-mediated mitophagy in Tibetan high-altitude adaptation

doi: 10.1038/s41420-025-02933-8

Figure Lengend Snippet: A Schematic diagram of the mechanism of action of PX-478. B Western blot analysis of HIF-1α, BNIP3, NIX, DRP1, LC3B, and p62 protein levels in M9a+sh EPAS1 cells following PX-478 treatment under hypoxia. Grayscale analysis of protein levels was normalized to β-Tubulin. C Autophagic flux was quantified by colocalizing LC3B (green) and Mitotracker (red) (mitophagosomes) using immunofluorescence in M9a+sh EPAS1 cells following PX-478 treatment under hypoxia. DAPI (blue) stains the nucleus. Scale bar: 20 μm. D – F Mitochondrial OCR (basal, ATP-linked, maximal), MMP, and ATP levels were determined in M9a+sh EPAS1 cells after PX-478 treatment under hypoxia. G CCK-8 assay showing cell viability in M9a+sh EPAS1 cells after PX-478 treatment under hypoxia. H Cell apoptosis was measured in M9a+sh EPAS1 cells after PX-478 treatment under hypoxia using Annexin V-FITC staining. I Western blot analysis of C-Caspase-3 and BAX protein levels in M9a+sh EPAS1 cells following PX-478 treatment under hypoxia. Grayscale analysis of protein levels was normalized to β-Tubulin. J Cytoplasmic free mtDNA was determined by staining dsDNA (green) and Mitotracker (red) in M9a+sh EPAS1 cells following PX-478 treatment under hypoxia. DAPI (blue) stains the nucleus. dsDNA that does not colocalize with either the nucleus or mitochondria is considered cytoplasmic free mtDNA. Scale bar: 20 μm. K Western blot analysis of NLRP3, IL1β, cGAS, and STING protein levels in M9a+sh EPAS1 cells after PX-478 treatment under hypoxia. Grayscale analysis of protein levels was normalized to β-Tubulin. L Inflammatory cytokines IL6, IL1β, and TNF-α in the culture medium were measured in M9a+sh EPAS1 cells after PX-478 treatment under hypoxia using an ELISA kit. M9a+sh EPAS1 cells ( n = 4) were treated with 30 μM PX-478 (for the last 22 h) under hypoxia (1% O₂) for 48 h. #1–4 represent individual samples. Data are presented as the mean ± SD of three independent experiments. Two-tailed, paired Student’s t tests; * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: Human IL-6, IL-1β, and TNF-α levels were measured using ELISA kits (LiankeBio, China) according to the manufacturer’s instructions.

Techniques: Western Blot, Immunofluorescence, CCK-8 Assay, Staining, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Cell Death Discovery

Article Title: Mitochondrial retrograde signaling initiates HIF-1α/BNIP3/NIX-mediated mitophagy in Tibetan high-altitude adaptation

doi: 10.1038/s41420-025-02933-8

Figure Lengend Snippet: A Schematic diagram of the mechanism of action of Mdivi-1. B Western blot analysis of DRP1, NIX, BNIP3, LC3B, and p62 protein levels in M9a+sh EPAS1 cells after Mdivi-1 treatment under hypoxia. Grayscale analysis of protein levels was normalized to β-Tubulin. C Autophagic flux was assessed by quantifying the colocalization of LC3B (green) and Mitotracker (red) (mitophagosomes) using immunofluorescence in M9a+sh EPAS1 cells after Mdivi-1 treatment under hypoxia. DAPI (blue) stains the nucleus. Scale bar: 20 μm. D – F Mitochondrial OCR (basal, ATP-linked, maximal), MMP, and ATP content were measured in M9a+sh EPAS1 cells after Mdivi-1 treatment under hypoxia. G CCK-8 assay showing cell viability in M9a+sh EPAS1 cells after Mdivi-1 treatment under hypoxia. H Cell apoptosis was determined in M9a+sh EPAS1 cells after Mdivi-1 treatment under hypoxia using Annexin V-FITC staining. I Western blot analysis of C-Caspase-3 and BAX protein levels in M9a+sh EPAS1 cells after Mdivi-1 treatment under hypoxia. Grayscale analysis of protein levels was normalized to β-Tubulin. J Cytoplasmic free mtDNA was determined by staining dsDNA (green) and Mitotracker (red) using immunofluorescence in M9a+sh EPAS1 cells after Mdivi-1 treatment under hypoxia. DAPI (blue) stains the nucleus. dsDNA that does not colocalize with either the nucleus or mitochondria is considered cytoplasmic free mtDNA. Scale bar: 20 μm. K Western blot analysis of NLRP3, IL1β, cGAS, and STING protein levels in M9a+sh EPAS1 cells after Mdivi-1 treatment under hypoxia. Grayscale analysis of protein levels was normalized to β-Tubulin. L Inflammatory cytokines IL6, IL1β, and TNF-α in the culture medium were measured using ELISA kits in M9a+sh EPAS1 cells after Mdivi-1 treatment under hypoxia. M9a+sh EPAS1 cells ( n = 4) were treated with 50 μM Mdivi-1 or DMSO (pre-treatment for 4 h, followed by treatment for 48 h) under hypoxia (1% O₂).#1–4 represent individual samples. Data are presented as the mean ± SD of three independent experiments. Two-tailed, paired Student’s t tests; P < 0.05; * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: Human IL-6, IL-1β, and TNF-α levels were measured using ELISA kits (LiankeBio, China) according to the manufacturer’s instructions.

Techniques: Western Blot, Immunofluorescence, CCK-8 Assay, Staining, Enzyme-linked Immunosorbent Assay, Two Tailed Test