Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: LncRNA SYISL promotes fibroblast myofibroblast transition via miR-23a-mediated TRIOBP regulation

doi: 10.1007/s00018-025-05729-2

Figure Lengend Snippet: Overexpression of SYISL significantly promotes the differentiation of pulmonary fibroblasts into myofibroblasts. A RT-qPCR analysis validated the mRNA levels of SYISL, Acta2, Fn1, and Col1a1 in PMLFs. Bars represent mean values of mRNA levels normalized to Gapdh mRNA. B Western blot assay showing the protein level of Fibronectin, α-sma, Collagen I, Triobp, and Gapdh in PMLFs. C Proliferative capacity was analyzed using an EDU assay in PMLFs. The nuclei were counterstained with DAPI. Bar = 50 μm. D SYISL is critical for cell metastasis in a transwell assay. Bar = 100 μm. E The activation of PMLFs was examined based on fibroblast contraction in 3D collagen matrices. F Immunofluorescence analysis revealed that SYISL significantly heightened the expression of α-sma in PMLFs. The nuclei were counterstained with DAPI. Bar = 20 μm. G RT-qPCR analysis validated the mRNA levels of ACTA2, FN1, and COL1 A1 in PHLFs. Bars represent mean values of mRNA levels normalized to GAPDH mRNA. H Western blot assay showing the protein level of Fibronectin, Collagen I, α-SMA, TRIOBP, and GAPDH in PHLFs. I Proliferative capacity was analyzed using an EDU assay in PHLFs. The nuclei were counterstained with DAPI. Bar = 50 μm. J Immunofluorescence analysis revealed that SYISL significantly heightened the expression of α-SMA in PHLFs. The nuclei were counterstained with DAPI. Bar = 20 μm. K The activation of PHLFs was examined based on fibroblast contraction in 3D collagen matrices. All Data are presented as mean ± SD (* p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001, unpaired Student’s t-test unpaired Student’s t-test)

Article Snippet: The following antibodies were used: TRIOBP (16124–1-AP, Proteintech), GAPDH (AF7021, Affinity), α-SMA (ab124964, Abcam), Type I collagen (14695–1-AP, Proteintech), Fibronectin (15613–1-AP, Proteintech), Vim (10366–1-AP, Proteintech).

Techniques: Over Expression, Quantitative RT-PCR, Western Blot, EdU Assay, Transwell Assay, Activation Assay, Immunofluorescence, Expressing

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: LncRNA SYISL promotes fibroblast myofibroblast transition via miR-23a-mediated TRIOBP regulation

doi: 10.1007/s00018-025-05729-2

Figure Lengend Snippet: SYISL bound and inhibited the expression of miR-23a to promote lung fibroblast activation. A Predicted binding sites of SYISL or hSYISL and miR-23a.delete, deleted binding site. B Luciferase reporter activities of chimeric vectors carrying the luciferase gene and a fragment of SYISL and hSYISL containing wild-type or deleted miR-23a-binding sites. **** p < 0.0001 when compared with the plasmid cotransfected with empty vector and TRIOBP‐wild-type; two-way ANOVA evaluated data with Šídák's multiple comparisons test for pairwise comparisons ( n = 3). C - D RT-qPCR analysis of expression of miR-23a after SYISL knockdown in PHLFs(C) and PMLFs(D). Bars represent mean values of RNA levels normalized to U6. E RT-qPCR analysis of expression of miR-23a after SYISL overexpression PMLFs, Bars represent mean values of RNA levels normalized to U6. F RT-qPCR analysis of expression of SYISL in PMLFs after transfection mimic miR-23a and inhibitor miR-23a.Bars represent mean values of mRNA levels normalized to Gapdh mRNA. G RT-qPCR analysis of miR-23a expression in peripheral blood samples from IPF patients and healthy controls. Bars represent mean values of mRNA levels normalized to U6. H RT-qPCR analysis of the mRNA levels of miR-23a, Fibronectin, COL1 A1, α-SMA in PHLFs. The PHLFs were transfected with mimic miR-23a for 24 h and then were treated with 10 ng/ml TGF-β for another 24 h. Bars represent mean values of mRNA levels normalized to GAPDH mRNA orU6. I Western blot assay showing the protein level of Fibronectin, Collagen I, TRIOBP, α-SMA, and GAPDH in PHLFs that were transfected with mimic miR-23a for 24 h, and then were treated with 10 ng/ml TGF-β for another 24 h. J Proliferative capacity was analyzed using an EDU assay in PHLFs. The PHLFs were transfected with mimic miR-23a for 24 h and then were treated with 10 ng/ml TGF-β for another 24 h. The nuclei were counterstained with DAPI. Bar = 50 μm. K The fibroblast-induced CGC assay demonstrated that miR-23a overexpression in PHLFs inhibited the fibroblast contractility. L Immunofluorescence staining of Vimentin in PHLFs. The nuclei were counterstained with DAPI. Bar = 20 μm. M Western blot assay showing the protein level of Fibronectin, TRIOBP, α-SMA, and GAPDH in PHLFs. All data are presented as mean ± SD (* p < 0.01, ** p < 0.01, *** p < 0.001 and **** p < 0.0001, unpaired Student’s t-test unpaired Student’s t-test)

Article Snippet: The following antibodies were used: TRIOBP (16124–1-AP, Proteintech), GAPDH (AF7021, Affinity), α-SMA (ab124964, Abcam), Type I collagen (14695–1-AP, Proteintech), Fibronectin (15613–1-AP, Proteintech), Vim (10366–1-AP, Proteintech).

Techniques: Expressing, Activation Assay, Binding Assay, Luciferase, Plasmid Preparation, Quantitative RT-PCR, Knockdown, Over Expression, Transfection, Western Blot, EdU Assay, Immunofluorescence, Staining

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: LncRNA SYISL promotes fibroblast myofibroblast transition via miR-23a-mediated TRIOBP regulation

doi: 10.1007/s00018-025-05729-2

Figure Lengend Snippet: MiR-23a inhibits lung fibroblast activation by repressing TRIOBP expression. A Predicted binding sites of TRIOBP or Triobp and miR-23a.delete, deleted binding site. B Luciferase reporter assay shows the interaction between miR-23a-3p and TRIOBP/Triobp-WT or TRIOBP/Triobp-delete (fragment with the 3, UTR containing the WT or delete-miR-23a-3p binding site). C - D RT-qPCR analysis showing that miR-23a expression after knockdown of TRIOBP in PHLFS cells (C) and PMLFs (D). Bars represent mean values of RNA levels normalized to U6. E RT-qPCR analysis validated the mRNA levels of Triobp after transfection mimic miR-23a and inhibitor miR-23a in PMLFs. Bars represent mean values of mRNA levels normalized to Gapdh mRNA. F Immunohistochemical analysis of lung tissue sections from patients with IPF revealed a pronounced upregulation of TRIOBP protein, specifically localized within the fibrotic foci. G - J RT-qPCR analysis of the mRNA levels of TRIOBP, ACTA2, COL1 A1, and FN1 in sh NC and shTRIOBP in PHLFs. The sh NC and shTIROBP in PHLFs were treated with 10 ng/ml TGF-β for 24 h. Bars represent mean values of mRNA levels normalized to GAPDH mRNA. K Western blot analysis showing the protein levels of TRIOBP, Fibronectin, α-SMA, Collagen I, and GAPDH in sh NC and shTRIOBP PHLFs cell lines after treatment with 10 ng/ml TGF-β for 24 h. L Proliferative capacity was analyzed using an EDU assay in PHLFs. The shNC and shTRIOBP in PHLFs were treated with 10 ng/ml TGF-β for 24 h. The nuclei were counterstained with DAPI. Bar = 20 μm. M The fibroblast-induced CGC assay demonstrated that the knockdown of TRIOBP in PHLFs inhibited the fibroblast contractility. N – O Western blot analysis showing the protein levels of Fibronectin, Collagen I, TRIOBP, α-SMA, and GAPDH in PHLFs transfected with mimic-23a (N) and inhibitor-23a (O) for 24 h, followed by treatment with 10 ng/ml TGF-β for an additional 24 h. All data are presented as mean ± SD (* p < 0.01, ** p < 0.01, *** p < 0.001 and **** p < 0.0001, unpaired Student’s t-test unpaired Student’s t-test)

Article Snippet: The following antibodies were used: TRIOBP (16124–1-AP, Proteintech), GAPDH (AF7021, Affinity), α-SMA (ab124964, Abcam), Type I collagen (14695–1-AP, Proteintech), Fibronectin (15613–1-AP, Proteintech), Vim (10366–1-AP, Proteintech).

Techniques: Activation Assay, Expressing, Binding Assay, Luciferase, Reporter Assay, Quantitative RT-PCR, Knockdown, Transfection, Immunohistochemical staining, Western Blot, EdU Assay

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: LncRNA SYISL promotes fibroblast myofibroblast transition via miR-23a-mediated TRIOBP regulation

doi: 10.1007/s00018-025-05729-2

Figure Lengend Snippet: SYISL Knockdown for the treatment of bleomycin-induced pulmonary fibrosis in mice. A Schematic representation of the experimental procedure. Wild-type (WT; C57BL/6 J) mice were administered with saline or BLM via tracheal intubation to establish a pulmonary fibrosis model, followed by injection of AAV-shNC and AAV-shSYISL adenoviruses seven days later, with tissue collection after 28 days. B RT-qPCR analysis validated that SYISL knockdown significantly reduced the expression of Acta2, Col1a1, and Fn1 mRNA. Bars represent the mean values of mRNA levels normalized to Gapdh mRNA. C Significantly reduced hydroxyproline content in the lung tissues of AAV6-shSYISL-treated mice, n = 10. Two-way ANOVA evaluated data with Šídák's multiple comparisons test for pairwise comparisons (**** p < 0.0001). D Western blot analysis of BLM-treated mice showed significantly reduced expression of Fn1, Col1a1, α-sma, and Triobp proteins following SYISL silencing; n = 3. E RT-qPCR analysis of the expression of miR-23a in the lungs of mice after injection of BLM with or without SYISL knockdown. Bars represent mean values of mRNA levels normalized to U6. F H&E and Mason’s trichrome staining from lung sections at day 28. IHC staining of Collagen I, Vimentin, α-sma and Triobp in the lungs of mice. Scale bars, 100 μm. All data are presented as mean ± SD (* p < 0.01, ** p < 0.01, *** p < 0.001 and **** p < 0.0001, unpaired Student’s t-test unpaired Student’s t-test)

Article Snippet: The following antibodies were used: TRIOBP (16124–1-AP, Proteintech), GAPDH (AF7021, Affinity), α-SMA (ab124964, Abcam), Type I collagen (14695–1-AP, Proteintech), Fibronectin (15613–1-AP, Proteintech), Vim (10366–1-AP, Proteintech).

Techniques: Knockdown, Saline, Injection, Quantitative RT-PCR, Expressing, Western Blot, Staining, Immunohistochemistry

Journal: World Journal of Gastroenterology

Article Title: Bletilla striata polysaccharides alleviate metabolic dysfunction-associated steatotic liver disease through enhancing hepatocyte RelA/ HNF1α signaling

doi: 10.3748/wjg.v31.i4.93179

Figure Lengend Snippet: Primary antibody information

Article Snippet: SREBP1 , Mouse mAb , Ma5-16124 , Thermo Fisher Scientific , Mouse, human.

Techniques: