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ha clover hif 1ɑ s31d  (Addgene inc)


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    Addgene inc ha clover hif 1ɑ s31d
    Fig. 8 HIF1ɑ mediates FBXL16 regulated fibroblast activation. (A) The mRNA expression of HIF1ɑ and FBXL16 in isolated primary human lung fibroblasts derived from COPD patients and normal counterparts (n = 6 per group). (B) MRC-5 were transfected with wild type and point mutated <t>(S31D)</t> HIF1ɑ and treated as indicated. The expression of HIF1ɑ was determined by western blotting. HA-tag was detected to confirm ectopic expression of HIF1ɑ. (C) The markers of fibroblast transdifferentiation from (B) were determined by real-time PCR (n = 3). The values were normalized by β-actin expression. (D) C57BL/6 mice were normally ventilated (Air) or exposed to chronic smoking (CS) for 16 weeks to induce experimental COPD. Mice were intranasally administered with control or FBXL16 expressing AAV6 twice. The protein abundance of HIF1ɑ and FBXL16 in the lungs of mice were measured. Left, the representative result was shown; right, the intensity of bands was calculated by ImageJ and normalized by β-actin expression (n = 3). (E) MRC-5 cells were transfected with HIF1ɑ and/or FBXL16 expressing plasmid and treated by TGF-β as indicated. The markers of lung fibroblast activation (ɑ-SMA and fibronetion) were determined. Left, the representative result was shown; right, the intensity of bands was calculated by ImageJ and normalized by β-actin expression (n = 3). (F) MRC-5 cells were transfected with control or miR-1307-5p agomir (miR) and treated by TGF-β as indicated. The protein abundance of HIF1ɑ was measured. Left, the representative result was shown; right, the intensity of bands was calculated by ImageJ and normalized by ɑ-tubulin expression (n = 3). (G) C57BL/6 mice were normally ventilated (Air) or exposed to chronic smoking (CS) for 16 weeks to induce experimental COPD. MiR- 1307-5p agomir (miR) or scrambled (Scr) agomir were treated intranasally weekly. The protein abundance of HIF1ɑ and FBXL16 in the lungs of mice were measured by western blotting. Left, the representative result was shown; right, the intensity of bands was calculated by ImageJ and normalized by β-actin expression (n = 3). Data are presented as means ± SD. n.s., not significant. *p < 0.05; **p < 0.01; ***p < 0.001
    Ha Clover Hif 1ɑ S31d, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ha clover hif 1ɑ s31d - by Bioz Stars, 2026-06
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    1) Product Images from "MiR-1307-5p enhances fibroblast transdifferentiation to exacerbate chronic obstructive pulmonary disease through regulating FBXL16/HIF1α axis."

    Article Title: MiR-1307-5p enhances fibroblast transdifferentiation to exacerbate chronic obstructive pulmonary disease through regulating FBXL16/HIF1α axis.

    Journal: Respiratory research

    doi: 10.1186/s12931-024-03007-6

    Fig. 8 HIF1ɑ mediates FBXL16 regulated fibroblast activation. (A) The mRNA expression of HIF1ɑ and FBXL16 in isolated primary human lung fibroblasts derived from COPD patients and normal counterparts (n = 6 per group). (B) MRC-5 were transfected with wild type and point mutated (S31D) HIF1ɑ and treated as indicated. The expression of HIF1ɑ was determined by western blotting. HA-tag was detected to confirm ectopic expression of HIF1ɑ. (C) The markers of fibroblast transdifferentiation from (B) were determined by real-time PCR (n = 3). The values were normalized by β-actin expression. (D) C57BL/6 mice were normally ventilated (Air) or exposed to chronic smoking (CS) for 16 weeks to induce experimental COPD. Mice were intranasally administered with control or FBXL16 expressing AAV6 twice. The protein abundance of HIF1ɑ and FBXL16 in the lungs of mice were measured. Left, the representative result was shown; right, the intensity of bands was calculated by ImageJ and normalized by β-actin expression (n = 3). (E) MRC-5 cells were transfected with HIF1ɑ and/or FBXL16 expressing plasmid and treated by TGF-β as indicated. The markers of lung fibroblast activation (ɑ-SMA and fibronetion) were determined. Left, the representative result was shown; right, the intensity of bands was calculated by ImageJ and normalized by β-actin expression (n = 3). (F) MRC-5 cells were transfected with control or miR-1307-5p agomir (miR) and treated by TGF-β as indicated. The protein abundance of HIF1ɑ was measured. Left, the representative result was shown; right, the intensity of bands was calculated by ImageJ and normalized by ɑ-tubulin expression (n = 3). (G) C57BL/6 mice were normally ventilated (Air) or exposed to chronic smoking (CS) for 16 weeks to induce experimental COPD. MiR- 1307-5p agomir (miR) or scrambled (Scr) agomir were treated intranasally weekly. The protein abundance of HIF1ɑ and FBXL16 in the lungs of mice were measured by western blotting. Left, the representative result was shown; right, the intensity of bands was calculated by ImageJ and normalized by β-actin expression (n = 3). Data are presented as means ± SD. n.s., not significant. *p < 0.05; **p < 0.01; ***p < 0.001
    Figure Legend Snippet: Fig. 8 HIF1ɑ mediates FBXL16 regulated fibroblast activation. (A) The mRNA expression of HIF1ɑ and FBXL16 in isolated primary human lung fibroblasts derived from COPD patients and normal counterparts (n = 6 per group). (B) MRC-5 were transfected with wild type and point mutated (S31D) HIF1ɑ and treated as indicated. The expression of HIF1ɑ was determined by western blotting. HA-tag was detected to confirm ectopic expression of HIF1ɑ. (C) The markers of fibroblast transdifferentiation from (B) were determined by real-time PCR (n = 3). The values were normalized by β-actin expression. (D) C57BL/6 mice were normally ventilated (Air) or exposed to chronic smoking (CS) for 16 weeks to induce experimental COPD. Mice were intranasally administered with control or FBXL16 expressing AAV6 twice. The protein abundance of HIF1ɑ and FBXL16 in the lungs of mice were measured. Left, the representative result was shown; right, the intensity of bands was calculated by ImageJ and normalized by β-actin expression (n = 3). (E) MRC-5 cells were transfected with HIF1ɑ and/or FBXL16 expressing plasmid and treated by TGF-β as indicated. The markers of lung fibroblast activation (ɑ-SMA and fibronetion) were determined. Left, the representative result was shown; right, the intensity of bands was calculated by ImageJ and normalized by β-actin expression (n = 3). (F) MRC-5 cells were transfected with control or miR-1307-5p agomir (miR) and treated by TGF-β as indicated. The protein abundance of HIF1ɑ was measured. Left, the representative result was shown; right, the intensity of bands was calculated by ImageJ and normalized by ɑ-tubulin expression (n = 3). (G) C57BL/6 mice were normally ventilated (Air) or exposed to chronic smoking (CS) for 16 weeks to induce experimental COPD. MiR- 1307-5p agomir (miR) or scrambled (Scr) agomir were treated intranasally weekly. The protein abundance of HIF1ɑ and FBXL16 in the lungs of mice were measured by western blotting. Left, the representative result was shown; right, the intensity of bands was calculated by ImageJ and normalized by β-actin expression (n = 3). Data are presented as means ± SD. n.s., not significant. *p < 0.05; **p < 0.01; ***p < 0.001

    Techniques Used: Activation Assay, Expressing, Isolation, Derivative Assay, Transfection, Western Blot, Real-time Polymerase Chain Reaction, Control, Quantitative Proteomics, Plasmid Preparation



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    Fig. 8 HIF1ɑ mediates FBXL16 regulated fibroblast activation. (A) The mRNA expression of HIF1ɑ and FBXL16 in isolated primary human lung fibroblasts derived from COPD patients and normal counterparts (n = 6 per group). (B) MRC-5 were transfected with wild type and point mutated <t>(S31D)</t> HIF1ɑ and treated as indicated. The expression of HIF1ɑ was determined by western blotting. HA-tag was detected to confirm ectopic expression of HIF1ɑ. (C) The markers of fibroblast transdifferentiation from (B) were determined by real-time PCR (n = 3). The values were normalized by β-actin expression. (D) C57BL/6 mice were normally ventilated (Air) or exposed to chronic smoking (CS) for 16 weeks to induce experimental COPD. Mice were intranasally administered with control or FBXL16 expressing AAV6 twice. The protein abundance of HIF1ɑ and FBXL16 in the lungs of mice were measured. Left, the representative result was shown; right, the intensity of bands was calculated by ImageJ and normalized by β-actin expression (n = 3). (E) MRC-5 cells were transfected with HIF1ɑ and/or FBXL16 expressing plasmid and treated by TGF-β as indicated. The markers of lung fibroblast activation (ɑ-SMA and fibronetion) were determined. Left, the representative result was shown; right, the intensity of bands was calculated by ImageJ and normalized by β-actin expression (n = 3). (F) MRC-5 cells were transfected with control or miR-1307-5p agomir (miR) and treated by TGF-β as indicated. The protein abundance of HIF1ɑ was measured. Left, the representative result was shown; right, the intensity of bands was calculated by ImageJ and normalized by ɑ-tubulin expression (n = 3). (G) C57BL/6 mice were normally ventilated (Air) or exposed to chronic smoking (CS) for 16 weeks to induce experimental COPD. MiR- 1307-5p agomir (miR) or scrambled (Scr) agomir were treated intranasally weekly. The protein abundance of HIF1ɑ and FBXL16 in the lungs of mice were measured by western blotting. Left, the representative result was shown; right, the intensity of bands was calculated by ImageJ and normalized by β-actin expression (n = 3). Data are presented as means ± SD. n.s., not significant. *p < 0.05; **p < 0.01; ***p < 0.001
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    Fig. 8 HIF1ɑ mediates FBXL16 regulated fibroblast activation. (A) The mRNA expression of HIF1ɑ and FBXL16 in isolated primary human lung fibroblasts derived from COPD patients and normal counterparts (n = 6 per group). (B) MRC-5 were transfected with wild type and point mutated <t>(S31D)</t> HIF1ɑ and treated as indicated. The expression of HIF1ɑ was determined by western blotting. HA-tag was detected to confirm ectopic expression of HIF1ɑ. (C) The markers of fibroblast transdifferentiation from (B) were determined by real-time PCR (n = 3). The values were normalized by β-actin expression. (D) C57BL/6 mice were normally ventilated (Air) or exposed to chronic smoking (CS) for 16 weeks to induce experimental COPD. Mice were intranasally administered with control or FBXL16 expressing AAV6 twice. The protein abundance of HIF1ɑ and FBXL16 in the lungs of mice were measured. Left, the representative result was shown; right, the intensity of bands was calculated by ImageJ and normalized by β-actin expression (n = 3). (E) MRC-5 cells were transfected with HIF1ɑ and/or FBXL16 expressing plasmid and treated by TGF-β as indicated. The markers of lung fibroblast activation (ɑ-SMA and fibronetion) were determined. Left, the representative result was shown; right, the intensity of bands was calculated by ImageJ and normalized by β-actin expression (n = 3). (F) MRC-5 cells were transfected with control or miR-1307-5p agomir (miR) and treated by TGF-β as indicated. The protein abundance of HIF1ɑ was measured. Left, the representative result was shown; right, the intensity of bands was calculated by ImageJ and normalized by ɑ-tubulin expression (n = 3). (G) C57BL/6 mice were normally ventilated (Air) or exposed to chronic smoking (CS) for 16 weeks to induce experimental COPD. MiR- 1307-5p agomir (miR) or scrambled (Scr) agomir were treated intranasally weekly. The protein abundance of HIF1ɑ and FBXL16 in the lungs of mice were measured by western blotting. Left, the representative result was shown; right, the intensity of bands was calculated by ImageJ and normalized by β-actin expression (n = 3). Data are presented as means ± SD. n.s., not significant. *p < 0.05; **p < 0.01; ***p < 0.001
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    Fig. 8 HIF1ɑ mediates FBXL16 regulated fibroblast activation. (A) The mRNA expression of HIF1ɑ and FBXL16 in isolated primary human lung fibroblasts derived from COPD patients and normal counterparts (n = 6 per group). (B) MRC-5 were transfected with wild type and point mutated <t>(S31D)</t> HIF1ɑ and treated as indicated. The expression of HIF1ɑ was determined by western blotting. HA-tag was detected to confirm ectopic expression of HIF1ɑ. (C) The markers of fibroblast transdifferentiation from (B) were determined by real-time PCR (n = 3). The values were normalized by β-actin expression. (D) C57BL/6 mice were normally ventilated (Air) or exposed to chronic smoking (CS) for 16 weeks to induce experimental COPD. Mice were intranasally administered with control or FBXL16 expressing AAV6 twice. The protein abundance of HIF1ɑ and FBXL16 in the lungs of mice were measured. Left, the representative result was shown; right, the intensity of bands was calculated by ImageJ and normalized by β-actin expression (n = 3). (E) MRC-5 cells were transfected with HIF1ɑ and/or FBXL16 expressing plasmid and treated by TGF-β as indicated. The markers of lung fibroblast activation (ɑ-SMA and fibronetion) were determined. Left, the representative result was shown; right, the intensity of bands was calculated by ImageJ and normalized by β-actin expression (n = 3). (F) MRC-5 cells were transfected with control or miR-1307-5p agomir (miR) and treated by TGF-β as indicated. The protein abundance of HIF1ɑ was measured. Left, the representative result was shown; right, the intensity of bands was calculated by ImageJ and normalized by ɑ-tubulin expression (n = 3). (G) C57BL/6 mice were normally ventilated (Air) or exposed to chronic smoking (CS) for 16 weeks to induce experimental COPD. MiR- 1307-5p agomir (miR) or scrambled (Scr) agomir were treated intranasally weekly. The protein abundance of HIF1ɑ and FBXL16 in the lungs of mice were measured by western blotting. Left, the representative result was shown; right, the intensity of bands was calculated by ImageJ and normalized by β-actin expression (n = 3). Data are presented as means ± SD. n.s., not significant. *p < 0.05; **p < 0.01; ***p < 0.001
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    Fig. 8 HIF1ɑ mediates FBXL16 regulated fibroblast activation. (A) The mRNA expression of HIF1ɑ and FBXL16 in isolated primary human lung fibroblasts derived from COPD patients and normal counterparts (n = 6 per group). (B) MRC-5 were transfected with wild type and point mutated (S31D) HIF1ɑ and treated as indicated. The expression of HIF1ɑ was determined by western blotting. HA-tag was detected to confirm ectopic expression of HIF1ɑ. (C) The markers of fibroblast transdifferentiation from (B) were determined by real-time PCR (n = 3). The values were normalized by β-actin expression. (D) C57BL/6 mice were normally ventilated (Air) or exposed to chronic smoking (CS) for 16 weeks to induce experimental COPD. Mice were intranasally administered with control or FBXL16 expressing AAV6 twice. The protein abundance of HIF1ɑ and FBXL16 in the lungs of mice were measured. Left, the representative result was shown; right, the intensity of bands was calculated by ImageJ and normalized by β-actin expression (n = 3). (E) MRC-5 cells were transfected with HIF1ɑ and/or FBXL16 expressing plasmid and treated by TGF-β as indicated. The markers of lung fibroblast activation (ɑ-SMA and fibronetion) were determined. Left, the representative result was shown; right, the intensity of bands was calculated by ImageJ and normalized by β-actin expression (n = 3). (F) MRC-5 cells were transfected with control or miR-1307-5p agomir (miR) and treated by TGF-β as indicated. The protein abundance of HIF1ɑ was measured. Left, the representative result was shown; right, the intensity of bands was calculated by ImageJ and normalized by ɑ-tubulin expression (n = 3). (G) C57BL/6 mice were normally ventilated (Air) or exposed to chronic smoking (CS) for 16 weeks to induce experimental COPD. MiR- 1307-5p agomir (miR) or scrambled (Scr) agomir were treated intranasally weekly. The protein abundance of HIF1ɑ and FBXL16 in the lungs of mice were measured by western blotting. Left, the representative result was shown; right, the intensity of bands was calculated by ImageJ and normalized by β-actin expression (n = 3). Data are presented as means ± SD. n.s., not significant. *p < 0.05; **p < 0.01; ***p < 0.001

    Journal: Respiratory research

    Article Title: MiR-1307-5p enhances fibroblast transdifferentiation to exacerbate chronic obstructive pulmonary disease through regulating FBXL16/HIF1α axis.

    doi: 10.1186/s12931-024-03007-6

    Figure Lengend Snippet: Fig. 8 HIF1ɑ mediates FBXL16 regulated fibroblast activation. (A) The mRNA expression of HIF1ɑ and FBXL16 in isolated primary human lung fibroblasts derived from COPD patients and normal counterparts (n = 6 per group). (B) MRC-5 were transfected with wild type and point mutated (S31D) HIF1ɑ and treated as indicated. The expression of HIF1ɑ was determined by western blotting. HA-tag was detected to confirm ectopic expression of HIF1ɑ. (C) The markers of fibroblast transdifferentiation from (B) were determined by real-time PCR (n = 3). The values were normalized by β-actin expression. (D) C57BL/6 mice were normally ventilated (Air) or exposed to chronic smoking (CS) for 16 weeks to induce experimental COPD. Mice were intranasally administered with control or FBXL16 expressing AAV6 twice. The protein abundance of HIF1ɑ and FBXL16 in the lungs of mice were measured. Left, the representative result was shown; right, the intensity of bands was calculated by ImageJ and normalized by β-actin expression (n = 3). (E) MRC-5 cells were transfected with HIF1ɑ and/or FBXL16 expressing plasmid and treated by TGF-β as indicated. The markers of lung fibroblast activation (ɑ-SMA and fibronetion) were determined. Left, the representative result was shown; right, the intensity of bands was calculated by ImageJ and normalized by β-actin expression (n = 3). (F) MRC-5 cells were transfected with control or miR-1307-5p agomir (miR) and treated by TGF-β as indicated. The protein abundance of HIF1ɑ was measured. Left, the representative result was shown; right, the intensity of bands was calculated by ImageJ and normalized by ɑ-tubulin expression (n = 3). (G) C57BL/6 mice were normally ventilated (Air) or exposed to chronic smoking (CS) for 16 weeks to induce experimental COPD. MiR- 1307-5p agomir (miR) or scrambled (Scr) agomir were treated intranasally weekly. The protein abundance of HIF1ɑ and FBXL16 in the lungs of mice were measured by western blotting. Left, the representative result was shown; right, the intensity of bands was calculated by ImageJ and normalized by β-actin expression (n = 3). Data are presented as means ± SD. n.s., not significant. *p < 0.05; **p < 0.01; ***p < 0.001

    Article Snippet: HA-CloverHIF-1α wild-type (#163365) and HA-Clover-HIF-1ɑ S31D (#163370) were ordered from Addgene.

    Techniques: Activation Assay, Expressing, Isolation, Derivative Assay, Transfection, Western Blot, Real-time Polymerase Chain Reaction, Control, Quantitative Proteomics, Plasmid Preparation