Journal: Pflugers Archiv : European journal of physiology

Article Title: The effect of chronic intermittent hypobaric hypoxia improving liver damage in metabolic syndrome rats through ferritinophagy.

doi: 10.1007/s00424-023-02860-6

Figure Lengend Snippet: Fig. 7 Effects of CIHH on the expression of fibrosis related protein of rat liver tissue. a Rep- resentative blots. b The effect of CIHH on the expression level of TGF-β1 and α-SMA. Values are mean ± SEM; n = 4 in each group. *P < 0.05 vs. CON, ##P < 0.01 vs. MS

Article Snippet: FTH1 antibody was purchased from Cell Signaling Technology (#3998, Danvers, MA, USA); TfR (NB100-92243), FPN1 (NBP1-21,502), and α-SMA (NBP2-33,006) 1 3 antibodies were purchased from Novus (Saint Charles, MO, USA); NCOA4 antibody was purchased from Biorbyt (orb540576, Cambridge, UK); xCT (ab175186) and GPX4 (ab125066) antibodies were purchased from Abcam (Cambridge, UK); GAPDH (AB0037) and β-actin (AY0573) antibodies were purchased from Abways Company (Shanghai, China); TGF-β1 antibody (abs130620) and goat anti-rabbit IgG-Alexa Fluor 488 (abs20025) were purchased from Aibixin Biotechnology (Shanghai, China); Cy3-labeled goat anti-mouse IgG was purchased from Shanghai Biyuntian Biotechnology (A0562, Shanghai, China).

Techniques: Expressing

Journal: Nature Communications

Article Title: CD248-targeted BBIR-T cell therapy against late-activated fibroblasts in cardiac repair after myocardial infarction

doi: 10.1038/s41467-025-56703-2

Figure Lengend Snippet: a UMAP clustering of 13441 cardiac interstitial cells (left) and fibroblasts (right) isolated from C57BL/6 J wild-type (WT) mice at 7 and 14 days post-sham and post-MI surgery. Each dot represents a single cell. The expression of known marker genes annotated cell type. b UMAP and violin plots depict Postn , Acta2 , and Comp expression in fibroblasts. c Triple IF staining for periostin (green) and αSMA (red) in the peri-infarct zone of the hearts of C57BL/6 J WT mice 0, 7, 14, 21, and 28 days after MI surgery. Nuclei were stained blue. Scale bar = 100 μm. Quantification of the number of periostin + αSMA + F-Myo and periostin + αSMA - F-Act in each high-power field (HPF). n = 5 mice at each time point after MI. d Flow cytometry of fibroblasts isolated from hearts of C57BL/6 J WT and Pdgfra-CRE:tdTomato mice. Total cardiac fibroblasts were lineage traced by pdgfrα. F-Myo and F-Act were marked with antibodies against periostin-AF488 and αSMA-AF647. Quantification of the pdgfrα + fibroblast ratio in live cells, periostin + fibroblasts ratio in pdgfrα + fibroblasts, periostin + αSMA + F-Myo ratio in pdgfrα + fibroblasts, and periostin + αSMA - F-Act ratio in pdgfrα + fibroblasts. n = 5 mice at each time point after MI. e Spatial distribution of the F-Act and F-Myo subpopulation in mouse heart sections from sham and 10 days post-MI conditions, visualized through the integration of scRNA-seq and stereo-seq methodologies. c, d Data are the mean ± SEM and p -values are displayed in the bar charts, one-way ANOVA followed by Tukey’s multiple comparisons tests. Source data are provided as a Source Data file.

Article Snippet: The antibodies used in this study included Periostin (Abcam, ab14041), αSMA-Alexa Fluor 647 (Novus, NBP2-34522), CD248-Alexa Fluor 647 (Santa Cruz, sc-377221), CD3-FITC (BioLegend, 317306 for human and 100204 for mouse), CD4-APC (BioLegend, 344614 for human and 100412 for mouse), CD8-PE (BioLegend, 344706 for human and 100708 for mouse), and goat anti-rabbit IgG (1:1000, Invitrogen, A11008 for Alexa Fluor 488).

Techniques: Isolation, Expressing, Marker, Staining, Flow Cytometry

Journal: Nature Communications

Article Title: CD248-targeted BBIR-T cell therapy against late-activated fibroblasts in cardiac repair after myocardial infarction

doi: 10.1038/s41467-025-56703-2

Figure Lengend Snippet: a Triple IF staining for periostin (green) and CD248 (red) in the peri-infarct zone of the hearts of C57BL/6J WT mice 0, 7, 14, 21, and 28 days after MI surgery. Nuclei were stained blue. Scale bar = 100 μm. Quantification of the periostin + CD248 + cell to DAPI ratio in each HPF. n = 5 mice at each time point after MI. Triple IF staining of periostin, αSMA, COMP (stained green), and CD248 (stained red) in the hearts of MI mouses ( b ) and a patient with ischemic cardiomyopathy ( c ). Nuclei were stained blue. Scale bar =100 μm. IF staining of mouse heart was repeated in heart sections from 5 independent mice, while IF of human heart was repeated 5 times in heart sections from one patient with ischemic cardiomyopathy, yielding similar results. d Flow cytometry of cells isolated from hearts of C57BL/6 J WT mouses and Pdgfra-CRE:tdTomato mice. Total cardiac fibroblasts were lineage traced by pdgfrα. Quantification of the CD248 + cell ratio in live cells, pdgfrα + fibroblast ratio in CD248 + cells, CD248 + cell ratio in pdgfrα + fibroblasts, periostin + CD248 + cell ratio in pdgfrα + fibroblasts, CD248 + cell ratio in periostin + activated fibroblasts, and periostin + activated fibroblasts in CD248 + cells. n = 5 mice at each time point after MI. a, d Data are the mean ± SEM and p -values are displayed in the bar charts, one-way ANOVA followed by Tukey’s multiple comparisons tests. Source data are provided as a Source Data file.

Article Snippet: The antibodies used in this study included Periostin (Abcam, ab14041), αSMA-Alexa Fluor 647 (Novus, NBP2-34522), CD248-Alexa Fluor 647 (Santa Cruz, sc-377221), CD3-FITC (BioLegend, 317306 for human and 100204 for mouse), CD4-APC (BioLegend, 344614 for human and 100412 for mouse), CD8-PE (BioLegend, 344706 for human and 100708 for mouse), and goat anti-rabbit IgG (1:1000, Invitrogen, A11008 for Alexa Fluor 488).

Techniques: Staining, Flow Cytometry, Isolation

Journal: Nature Communications

Article Title: CD248-targeted BBIR-T cell therapy against late-activated fibroblasts in cardiac repair after myocardial infarction

doi: 10.1038/s41467-025-56703-2

Figure Lengend Snippet: a Schematic diagram of the structure of the BBIR-T system. Created in BioRender. Haiting, C. (2025) https://BioRender.com/l46n352 . b Schematic images of dcAv. BBIR-28-137ζ and CD19. CAR-137ζ sequence. c Western blotting shows exogenous CD3ζ expression in HEK-293T cells transduced with lentivirus (LV)-dcAv.BBIR and LV-anti-CD19.CAR. The samples derive from the same experiment and that gels/blots were processed in parallel. Quantifying the relative protein expression of CD3ζ to GAPDH. n = 4 cell culture wells of each group. d Flow cytometry of human peripheral T cells transduced with LV-dcAv.BBIR and LV-anti-CD19.CAR. Quantifying GFP + cells ratio to total T cells and biotin + cells ratio to GFP + cells. n = 5 cell culture wells of each group. e Western blotting by BF shows CD248 expression in NIH-3T3 cells with or without Cd248 overexpression(OE) or knockout(KO). The samples were derived from the same experiment and that gels/blots were processed in parallel. Quantifying the relative protein expression of CD248 to GAPDH. n = 4 cell culture wells of each group. f Triple IF staining of periostin, αSMA, COMP (green), and biotinylated anti-CD248 F(ab’)2 (shorted as BF) (red) in the peri-infarct zone 14 days after MI surgery. Nuclei were stained blue. Scale bar =100 μm. Each experiment was repeated 5 times independently with similar results. c-e Data are the mean ± SEM and p -values are displayed in the bar charts, one-way ANOVA followed by Tukey’s multiple comparisons tests. Source data are provided as a Source Data file.

Article Snippet: The antibodies used in this study included Periostin (Abcam, ab14041), αSMA-Alexa Fluor 647 (Novus, NBP2-34522), CD248-Alexa Fluor 647 (Santa Cruz, sc-377221), CD3-FITC (BioLegend, 317306 for human and 100204 for mouse), CD4-APC (BioLegend, 344614 for human and 100412 for mouse), CD8-PE (BioLegend, 344706 for human and 100708 for mouse), and goat anti-rabbit IgG (1:1000, Invitrogen, A11008 for Alexa Fluor 488).

Techniques: Sequencing, Western Blot, Expressing, Transduction, Cell Culture, Flow Cytometry, Over Expression, Knock-Out, Derivative Assay, Staining

Journal: Neurotrauma Reports

Article Title: Early Transcutaneous Tibial Nerve Stimulation Acutely Improves Lower Urinary Tract Function in Spinal Cord Injured Rats

doi: 10.1089/neur.2021.0058

Figure Lengend Snippet: Assessment of bladder hypertrophy four weeks after spinal cord injury. ( A ) Bladder weight. ( B ) Quantification of the muscle to collagen ratio obtained by Masson Trichrome staining. ( C ) Representative Masson Trichrome staining of bladder tissue. Muscle is stained red, collagen blue. ( D–F ) Quantification of three muscle proteins by Western blot (WES): myosin heavy chain 11 (MYH11; 182 kDa), alpha-smooth muscle actin (SMA; 48 kDa), and calponin (45 kDa) in sham and stimulated groups. GAPDH, glyceraldehyde 3-phosphate dehydrogenase. * p < 0.05.

Article Snippet: The primary antibodies used for detection were mouse primary antibodies against alpha smooth muscle actin (αSMA) (Novus Biologicals, Centennial, CO, NBP2-33006, 1:100), calponin (Sigma-Aldrich, C2687, 1:400), and myosin heavy chain 11 (MYH11) (Santa Cruz, sc-6956, 1:10).

Techniques: Staining, Western Blot

Journal: Respiratory Research

Article Title: Effect of active vitamin D3 on VEGF-induced ADAM33 expression and proliferation in human airway smooth muscle cells: implications for asthma treatment

doi: 10.1186/s12931-016-0490-9

Figure Lengend Snippet: 1,25(OH)2D3 inhibits VEGF-induced ADAM33 expression at both mRNA and protein level. ASM cells were incubated with various doses of 1,25(OH)2D3 for 9 h before treatment or not with 50 ng/ml of VEGF for 30 min, and then real-time PCR performed ( a ). ASM cells were incubated at indicated times of 100 nM of 1,25(OH)2D3, and then real-time PCR performed ( b ). The values are normalized relative to the GAPDH standard. ASM cells were incubated with various doses of 1,25(OH)2D3 for 24 h before treatment or not with 50 ng/ml of VEGF for 30 min, and then western blotting analysis for ADAM33 was performed ( c ). ASM cells were incubated at indicated times of 100 nM of 1,25(OH)2D3 before treatment or not with 50 ng/ml of VEGF for 30 min, and then western blotting analysis for ADAM33 was performed ( d ). β-actin was used as a loading control. All data are representative of three independent experiments. Values represent the means ± SEM. * P < 0.05 vs. control, # P < 0.05, ## P < 0.005 vs. VEGF alone

Article Snippet: ADAM33 was transfected into ASM cells, ASM cells-ADAM33, and ASM cells-vector according to a siRNA transfection protocol provided by Santa Cruz Biotechnology.

Techniques: Expressing, Incubation, Real-time Polymerase Chain Reaction, Western Blot, Control

Journal: Respiratory Research

Article Title: Effect of active vitamin D3 on VEGF-induced ADAM33 expression and proliferation in human airway smooth muscle cells: implications for asthma treatment

doi: 10.1186/s12931-016-0490-9

Figure Lengend Snippet: 1,25(OH)2D3 inhibits cell proliferation by down-regulation of ADAM33 expression. ASM cells were incubated with various doses of 1,25(OH)2D3 for 48 h before treatment or not with 50 ng/ml of VEGF for 30 min, and then cell proliferation was determined by BrdU incorporation ( a ). ASM cells were incubated at indicated times of 100 nM of 1,25(OH)2D3 before treatment or not with 50 ng/ml of VEGF for 30 min, and then cell proliferation was determined by BrdU incorporation ( b ). ASM cells were transfected with negative siRNA or ADAM33 siRNA, and then real-time PCR performed. The values are normalized relative to the GAPDH standard ( c ). ASM cells ( d ) and ASM cells-ADAM33 ( e ) were transfected with negative siRNA or ADAM33 siRNA, and then western blotting analysis for ADAM33 was performed. β-actin was used as a loading control. ASM cells-ADAM33 were transfected with negative siRNA or ADAM33 siRNA in the presence of VEGF (50 ng/ml) and 1,25-(OH)2D3 (100 nM) for 48 h, and then cell proliferation was determined by BrdU incorporation ( f ). All experiments were done at least three times. Values represent the means ± SEM. * P < 0.05 vs. control or ASMs-vector; # P < 0.05 vs. VEGF alone or control siRNA or ASMs-control siRNA

Article Snippet: ADAM33 was transfected into ASM cells, ASM cells-ADAM33, and ASM cells-vector according to a siRNA transfection protocol provided by Santa Cruz Biotechnology.

Techniques: Expressing, Incubation, BrdU Incorporation Assay, Transfection, Real-time Polymerase Chain Reaction, Western Blot, Control, Plasmid Preparation

Journal: Respiratory Research

Article Title: Effect of active vitamin D3 on VEGF-induced ADAM33 expression and proliferation in human airway smooth muscle cells: implications for asthma treatment

doi: 10.1186/s12931-016-0490-9

Figure Lengend Snippet: 1,25(OH)2D3 inhibits VEGF-induced ADAM33 expression and cell proliferation by inactivation of VEGFR2. ASM cells were incubated with indicated doses of SU1498 for 2 h before treatment with VEGF (50 ng/ml) for 24 h, and then western blotting analysis for ADAM33 was performed. β-actin was used as a loading control ( a ). ASM cells were incubated with indicated doses of SU1498 for 2 h before treatment with VEGF (50 ng/ml) for 48 h, and then cell proliferation was determined by BrdU incorporation ( b ). ASM cells were incubated with various doses of 1,25(OH)2D3 for 24 h before treatment or not with 50 ng/ml of VEGF for 30 min, and then western blotting analysis for VEGFR2 was performed ( c ). All experiments were done at least three times. Values represent the means ± SEM. * P < 0.05 vs. control; # P < 0.05, ## P < 0.005 vs. VEGF alone

Article Snippet: ADAM33 was transfected into ASM cells, ASM cells-ADAM33, and ASM cells-vector according to a siRNA transfection protocol provided by Santa Cruz Biotechnology.

Techniques: Expressing, Incubation, Western Blot, Control, BrdU Incorporation Assay

Journal: Respiratory Research

Article Title: Effect of active vitamin D3 on VEGF-induced ADAM33 expression and proliferation in human airway smooth muscle cells: implications for asthma treatment

doi: 10.1186/s12931-016-0490-9

Figure Lengend Snippet: 1,25(OH)2D3 inhibits VEGF-induced ERK 1/2 phosphorylation in ASM cells. ASM cells were incubated at indicated times of VEGF (50 ng/ml), and then western blotting analysis for phospho-ERK 1/2 ( a ) and phospho-Akt ( b ) was performed. ASM cells were incubated with various doses of 1,25(OH)2D3 for 24 h before treatment or not with 50 ng/ml of VEGF for 30 min, and then western blotting analysis for phospho-ERK 1/2 ( c ) and phospho-Akt ( d ) was performed. The total ERK1/2 and Akt was used as a loading control. ASM cells were incubated with 20 μM U0126 or 20 μM LY294002 for 2 h before treatment with VEGF (50 ng/ml) for 24 h, and then western blotting analysis for ADAM33 was performed. β-actin was used as a loading control ( e ). ASM cells were incubated with 20 μM U0126 or 20 μM LY294002 for 2 h before treatment with VEGF (50 ng/ml) for 48 h, and then cell proliferation was determined by BrdU incorporation ( f ). All experiments were done at least three times. Values represent the means ± SEM. * P < 0.05 vs. control; # P < 0.05 vs. VEGF alone

Article Snippet: ADAM33 was transfected into ASM cells, ASM cells-ADAM33, and ASM cells-vector according to a siRNA transfection protocol provided by Santa Cruz Biotechnology.

Techniques: Phospho-proteomics, Incubation, Western Blot, Control, BrdU Incorporation Assay