Journal: Scientific Reports

Article Title: Follistatin like-1 aggravates silica-induced mouse lung injury

doi: 10.1038/s41598-017-00478-0

Figure Lengend Snippet: Up-regulation of Fstl1 in silica-injured mice and patients with silicosis. ( a ) Fstl1 mRNA expression in lung tissues of C57BL/6J mice at the different time-point after silica injury was determined by qRT-PCR analysis (n = 6 per group; * P < 0.05; ** P < 0.01 by one-way ANOVA followed by Student’s t test). ( b ) FSTL1 protein in lung tissues of C57BL/6J mice at the different time-point after silica injury was determined by western blot analysis. β-tubulin was used as a loading control. ( c ) Immunohistochemistry (IHC) of FSTL1 in lung sections of C57BL/6J mice 21 days after saline or silica injury. Representative images of the staining are shown. NA stands for normal area, FA stands for fibrotic area; both are shown at higher magnification. (n = 6 per group; scale bars, 200 μm). ( d ) Fstl1 mRNA expression in primary alveolar macrophages (AMs), alveolar epithelial cells (AECs) and fibroblasts (Fb) at day 21 after saline or silica injury was determined by qRT-PCR analysis (n = 6 per group; * P < 0.05 by one-way ANOVA followed by Student’s t test). ( e ) FSTL1 protein in serum of C57BL/6J mice at the different time-point after silica injury was determined by ELISA analysis (n = 6 per group; * P < 0.05; ** P < 0.01 by one-way ANOVA followed by Student’s t test). ( f ) Representative images of lung fibrotic area of patient with silicosis with H&E staining and immunohistochemical staining for FSTL1 protein (NA stands for normal area, FA stands for fibrotic area; Scale bars, 200 μm). ( g ) FSTL1 levels in serum of patients with silicosis and normal control individuals were determined by ELISA (* P < 0.05 by one-way ANOVA followed by Student’s t test).

Article Snippet: The plate was incubated at 4 °C overnight, then 2.5 μg/mL biotinylated human FSTL1 detection antibody (R&D system, MAB1694) was added followed by washing with wash buffer and the plate was incubated for 1 hour at room temperature.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Control, Immunohistochemistry, Saline, Staining, Enzyme-linked Immunosorbent Assay, Immunohistochemical staining

Journal: Scientific Reports

Article Title: Follistatin like-1 aggravates silica-induced mouse lung injury

doi: 10.1038/s41598-017-00478-0

Figure Lengend Snippet: Fstl1 +/− mice have an attenuated pulmonary fibrosis phenotype. ( a ) Western blot analysis of FSTL1 expression in lung tissues of Fstl1 +/− and WT mice 21 days after saline or silica injury. β-tubulin was used as a loading control. ( b ) H&E staining of lung sections of Fstl1 +/− and WT mice 21 days after silica injury. Representative images of the staining are shown. Arrows show inflammatory cells. (n = 6 per group; scale bars, 200 μm). ( c ) Lung fibrotic score analysis of the lung sections from Fstl1 +/− and WT mice 21 days after silica injury (n = 6 per group; * P < 0.05 by one-way ANOVA followed by Student’s t test). The fibrotic area is presented as a percentage. ( d ) Hydroxyproline contents in lung tissues from Fstl1 +/− and WT mice were measured 21 days after saline or silica injury (n = 6 per group; * P < 0.05 by one-way ANOVA followed by Student’s t test). ( e ) Masson trichrome staining of lung sections of Fstl1 +/− and WT mice 21 days after silica treatment. Representative images of the staining are shown (n = 6 per group; scale bars, 200 μm). ( f ) qRT-PCR analysis of Col1a1 mRNA expression in lung tissues from Fstl1 +/− and WT mice 21 days after saline or silica injury (n = 3 per group; *** P < 0.001 by one-way ANOVA followed by Student’s t test). ( g ) Western blot analysis of type I collagen (Col1) expression in lung tissues from Fstl1 +/− and WT mice 21 days after saline or silica injury (n = 3 per group). β-tubulin was used as a loading control. ( h ) qRT-PCR analysis of fibronectin ( Fn1 ) mRNA expression in lung tissues from Fstl1 +/− and WT mice 21 days after saline or silica injury (n = 3 per group; * P < 0.05 by one-way ANOVA followed by Student’s t test).

Article Snippet: The plate was incubated at 4 °C overnight, then 2.5 μg/mL biotinylated human FSTL1 detection antibody (R&D system, MAB1694) was added followed by washing with wash buffer and the plate was incubated for 1 hour at room temperature.

Techniques: Western Blot, Expressing, Saline, Control, Staining, Quantitative RT-PCR

Journal: Scientific Reports

Article Title: Follistatin like-1 aggravates silica-induced mouse lung injury

doi: 10.1038/s41598-017-00478-0

Figure Lengend Snippet: Fstl1 +/− mice have an attenuated pulmonary inflammatory response. ( a–e ) Fstl1 +/− and their WT littermate mice were intratracheally exposured to saline or 200 mg/Kg silica, bronchoalveolar lavage fluid (BALF) were collected from Fstl1 +/− and WT mice 7 days after administration of saline or silica. ( a ) The number of total BALF cells was determined by hemocytometer (n = 7 per group; * P < 0.05 by one-way ANOVA followed by Student’s t test). ( b–d ) The differential cell counts in BALF were determined according to standard morphologic criteria. ( b ) Macrophages (n = 7 per group; * P < 0.05 by one-way ANOVA followed by Student’s t test). ( c ) PMNs (Polymorphonuclear neutrophils, n = 7 per group). ( d ) Lymphocytes (n = 7 per group). ( e ) The level of cytokine IL-1β in BALF was detected by ELISA assay (n = 7 per group; * P < 0.05 by one-way ANOVA followed by Student’s t test). ( f–g ) Fstl1 +/− and their WT littermate mice were intratracheally exposured to saline or silica, lung tissues were collected from Fstl1 +/− and WT mice 7 days after administration of saline or silica. ( f ) The level of cytokine IL-1β in lung tissues was detected by ELISA assay (n = 4 per group; * P < 0.05 by one-way ANOVA followed by Student’s t test). ( g ) The levels of NLRP3 and caspase-1 (p20) in lung tissues were determined by western blot analysis. β-tubulin was used as a loading control.

Article Snippet: The plate was incubated at 4 °C overnight, then 2.5 μg/mL biotinylated human FSTL1 detection antibody (R&D system, MAB1694) was added followed by washing with wash buffer and the plate was incubated for 1 hour at room temperature.

Techniques: Saline, Enzyme-linked Immunosorbent Assay, Western Blot, Control

Journal: Scientific Reports

Article Title: Follistatin like-1 aggravates silica-induced mouse lung injury

doi: 10.1038/s41598-017-00478-0

Figure Lengend Snippet: Fstl1 +/− mice have less myofibroblast accumulation after silica exposure. ( a ) Immunofluorescence analysis of α-SMA expression in lung sections of Fstl1 +/− and WT mice 21 days after saline or silica exposure. Representative images of the staining are shown. (α-SMA, green; Endomucin, red; nucleus, blue; scale bars, 200 μm). ( b ) qRT-PCR analysis of α-SMA mRNA expression in lung tissues from Fstl1 +/− and WT mice 21 days after saline or silica treatment (n = 3 per group; * P < 0.05 by one-way ANOVA followed by Student’s t test). ( c ) Western blot analysis of α-SMA expression in lung tissues from Fstl1 +/− and WT mice 21 days after saline or silica treatment. β-tubulin was used as a loading control.

Article Snippet: The plate was incubated at 4 °C overnight, then 2.5 μg/mL biotinylated human FSTL1 detection antibody (R&D system, MAB1694) was added followed by washing with wash buffer and the plate was incubated for 1 hour at room temperature.

Techniques: Immunofluorescence, Expressing, Saline, Staining, Quantitative RT-PCR, Western Blot, Control

Journal: Scientific Reports

Article Title: Follistatin like-1 aggravates silica-induced mouse lung injury

doi: 10.1038/s41598-017-00478-0

Figure Lengend Snippet: Fstl1 modulates myofibroblast differentiation via facilitating TGF-β1 signaling. ( a ) qRT-PCR analysis of TGF-β1 mRNA expression in lung tissues of Fstl1 +/− and WT mice at indicated time after saline or silica exposure (n = 4 per group; * P < 0.05 by one-way ANOVA followed by Student’s t test). ( b ) ELISA analysis of active form of TGF-β1 protein in lung tissues of Fstl1 +/− and WT mice at indicated time after saline or silica exposure (n = 4 per group; * P < 0.05 by one-way ANOVA followed by Student’s t test). ( c ) The levels of phosphorylation of Smad2/3 (p-Smad2/3), total Smad2/3 (T-Smad2/3), phosphorylation of Smad1/5 (p-Smad1/5) and total Smad1/5 (T-Smad1/5) in lung tissues of of Fstl1 +/− and WT mice 21 days after saline or silica exposure were determined by western blot analysis. β-tubulin was used as a loading control. ( d ) Primary lung fibroblasts from Fstl1 +/− and WT mice were treated with 5 ng/ml TGF-β1. Protein expressions of α-SMA in cell extracts and type I collagen (Col1) in medium 24 h after TGF-β1 treatment were determined by western blot analysis. β-tubulin was used as a loading control.

Article Snippet: The plate was incubated at 4 °C overnight, then 2.5 μg/mL biotinylated human FSTL1 detection antibody (R&D system, MAB1694) was added followed by washing with wash buffer and the plate was incubated for 1 hour at room temperature.

Techniques: Quantitative RT-PCR, Expressing, Saline, Enzyme-linked Immunosorbent Assay, Phospho-proteomics, Western Blot, Control

Journal: Scientific Reports

Article Title: Follistatin like-1 aggravates silica-induced mouse lung injury

doi: 10.1038/s41598-017-00478-0

Figure Lengend Snippet: FSTL1-neutralizing antibody attenuates silica-induced lung inflammation and subsequent pulmonary fibrosis in mice. ( a ) In a FSTL1 blockage experiment, C57BL/6 mice were intraperitoneally injected with 22B6 mAb or IgG1 (n = 6 per group) every other day from 1 day after silica challenge till the mice were sacrificed on day 7 for inflammation analysis or day 21 for fibrosis analysis. ( b–f ) For inflammation analysis, ( b ) the number of total BALF cells was determined by hemocytometer (* P < 0.05 by one-way ANOVA followed by Student’s t test). ( c–e ) The differential cell counts in BALF were determined according to standard morphologic criteria. ( c ) Macrophages (* P < 0.05 by one-way ANOVA followed by Student’s t test). ( d ) PMNs. ( e ) Lymphocytes. ( f ) The level of cytokine IL-1β was detected by ELISA assay (* P < 0.05 by one-way ANOVA followed by Student’s t test). ( g – i ) For fibrosis analysis, ( g ) hydroxyproline contents in lung tissues were measured (* P < 0.05, *** P < 0.001 by one-way ANOVA followed by Student’s t test). ( h ) Representative images of the H&E staining of lung sections are shown (Scale bars, 200 μm). ( i ) Lung fibrotic score analysis of the lung sections. The fibrotic area is presented as a percentage (* P < 0.05 by one-way ANOVA followed by Student’s t test). ( j ) Interventional dosing regimen of lung fibrosis model. C57BL/6J mice were intraperitoneally injected with 22B6 mAb or IgG1 (n = 10 per group) at indicated time after silica exposure, and lungs were harvested on day 21. ( k ) Representative images of the H&E staining of lung sections are shown (Scale bars, 200 μm). ( l ) Lung fibrotic score analysis of the lung sections. The fibrotic area is presented as a percentage (* P < 0.05 by one-way ANOVA followed by Student’s t test). ( m ) Hydroxyproline contents in lung tissues (*** P < 0.001 by one-way ANOVA followed by Student’s t test).

Article Snippet: The plate was incubated at 4 °C overnight, then 2.5 μg/mL biotinylated human FSTL1 detection antibody (R&D system, MAB1694) was added followed by washing with wash buffer and the plate was incubated for 1 hour at room temperature.

Techniques: Injection, Enzyme-linked Immunosorbent Assay, Staining

Journal: Scientific Reports

Article Title: Follistatin like-1 aggravates silica-induced mouse lung injury

doi: 10.1038/s41598-017-00478-0

Figure Lengend Snippet: Clinical characteristics of the study groups.

Article Snippet: The plate was incubated at 4 °C overnight, then 2.5 μg/mL biotinylated human FSTL1 detection antibody (R&D system, MAB1694) was added followed by washing with wash buffer and the plate was incubated for 1 hour at room temperature.

Techniques:

Journal: Immunology and cell biology

Article Title: Follistatin-like protein 1 modulates IL-17 signaling via IL-17RC regulation in stromal cells.

doi: 10.1038/icb.2017.26

Figure Lengend Snippet: Figure 1 FSTL-1 suppression by shRNA reduces IL-17/TNFα-mediated cytokine production. In the ST2 cell line, (a) Fstl1 transcript abundance and (b) FSTL-1 protein production increased following IL-17/TNFα stimulation but was reduced in cells transfected with shRNA targeting Fstl1 (gray bars) compared with control shRNA (black bars). IL-17/TNFα-dependent cytokine production is reduced by Fstl1 inhibition as determined by (c, e) transcript induction and (d, f) protein secretion. *Po0.05; **Po0.01; ***Po0.001; ****Po0.0001; NS, not significant.

Article Snippet: Il17rc and Fstl1 plasmid transfection Murine Il17rc (Addgene 46864, Cambridge, MA, USA), Fstl1 (Origene MR204305, Rockville, MD, USA) and empty vector pCMV6-Entry control (Origene PS 100001) plasmids were transformed in and grown in Mix and Go E. coli (Zymo Research, Irvine, CA, USA) in 5 ml Luria-Bertani broth containing selection antibiotic and prepared using the QIAprep Spin Miniprep Kit (Qiagen 27106), per the manufacturer’s instructions.

Techniques: shRNA, Transfection, Control, Inhibition

Journal: Immunology and cell biology

Article Title: Follistatin-like protein 1 modulates IL-17 signaling via IL-17RC regulation in stromal cells.

doi: 10.1038/icb.2017.26

Figure Lengend Snippet: Figure 2 FSTL-1 suppression by gene deletion reduces IL-17/TNFα-mediated cytokine production. In primary BMSCs, (a) Fstl1 transcript abundance and (b) FSTL-1 protein production increased following IL-17A and TNFα stimulation but was reduced in cells from FSTL-1 KO mice (gray bars) compared with WT littermate control mice (black bars). IL-17/TNFα-dependent cytokine production is reduced by Fstl1 inhibition as determined by (c, e) transcript induction and (d, f) protein secretion. *Po0.05, **Po0.01, ***Po0.001,****Po0.0001.

Article Snippet: Il17rc and Fstl1 plasmid transfection Murine Il17rc (Addgene 46864, Cambridge, MA, USA), Fstl1 (Origene MR204305, Rockville, MD, USA) and empty vector pCMV6-Entry control (Origene PS 100001) plasmids were transformed in and grown in Mix and Go E. coli (Zymo Research, Irvine, CA, USA) in 5 ml Luria-Bertani broth containing selection antibiotic and prepared using the QIAprep Spin Miniprep Kit (Qiagen 27106), per the manufacturer’s instructions.

Techniques: Control, Inhibition

Journal: Immunology and cell biology

Article Title: Follistatin-like protein 1 modulates IL-17 signaling via IL-17RC regulation in stromal cells.

doi: 10.1038/icb.2017.26

Figure Lengend Snippet: Figure 6 Ectopic Fstl1 expression rescues Il17rc expression and cytokine production in FSTL-1 KO cells. FSTL-1 KO BMSC transfection with pCMV-fst17 rescues (a) Fstl1 and (b) Il17rc transcript abundance. pCMV-Fstl1 complementation in FSTL-1 KO BMSCs rescues IL-17A/TNF-stimulated transcript levels of (c) Il6 and (d) Csf3. *Po0.05, **Po0.01.

Article Snippet: Il17rc and Fstl1 plasmid transfection Murine Il17rc (Addgene 46864, Cambridge, MA, USA), Fstl1 (Origene MR204305, Rockville, MD, USA) and empty vector pCMV6-Entry control (Origene PS 100001) plasmids were transformed in and grown in Mix and Go E. coli (Zymo Research, Irvine, CA, USA) in 5 ml Luria-Bertani broth containing selection antibiotic and prepared using the QIAprep Spin Miniprep Kit (Qiagen 27106), per the manufacturer’s instructions.

Techniques: Expressing, Transfection

Journal: Immunology and cell biology

Article Title: Follistatin-like protein 1 modulates IL-17 signaling via IL-17RC regulation in stromal cells.

doi: 10.1038/icb.2017.26

Figure Lengend Snippet: Figure 7 FSTL-1 influences Il17rc transcript abundance via transcriptional regulation. Following ActinomycinD treatment, FSTL-1 KO cells had reduced (a) Fstl1 and (b) Il17rc transcript abundance before ActinomycinD treatment and at 1, 2, 4 and 6 h posttreatment compared with WT BMSCs. Linear regression of Il17rc transcript abundance (c) at various time points following ActinomycinD treatment showed similar slopes for WT and FSTL-1 KO BMSCs (−3.254 ± 1.200 and −2.457 ± 1.429, P = 0.6748). Newly synthesized mRNA (5-EU labeled) was similar for (d) Hprt, but reduced for (e) Fstl1 and (f) Il17rc, transcripts in FSTL-1 KO cells compared with WT. *Po0.05, **Po0.01, ***Po0.001, ****Po0.0001.

Article Snippet: Il17rc and Fstl1 plasmid transfection Murine Il17rc (Addgene 46864, Cambridge, MA, USA), Fstl1 (Origene MR204305, Rockville, MD, USA) and empty vector pCMV6-Entry control (Origene PS 100001) plasmids were transformed in and grown in Mix and Go E. coli (Zymo Research, Irvine, CA, USA) in 5 ml Luria-Bertani broth containing selection antibiotic and prepared using the QIAprep Spin Miniprep Kit (Qiagen 27106), per the manufacturer’s instructions.

Techniques: Synthesized, Labeling

Journal: Immunology and cell biology

Article Title: Follistatin-like protein 1 modulates IL-17 signaling via IL-17RC regulation in stromal cells.

doi: 10.1038/icb.2017.26

Figure Lengend Snippet: Figure 8 Fstl1 expression correlates with Il17rc expression in the CD45-negative bone marrow population. CD45-negative cells from WT and FSTL-1 Hypomorphic mouse bone marrow had slightly reduced (a) Fstl1, (b) Il17rc and (c) Il17ra expression. Correlation analysis revealed highly significant association between (d) Fstl1 and Il17rc (r = 0.9512, R2 = 0.9048, Po0.0001) but not between (e) Fstl1 and Il17ra (r = −0.1982, R2 = 0.0393, P = 0.5162).

Article Snippet: Il17rc and Fstl1 plasmid transfection Murine Il17rc (Addgene 46864, Cambridge, MA, USA), Fstl1 (Origene MR204305, Rockville, MD, USA) and empty vector pCMV6-Entry control (Origene PS 100001) plasmids were transformed in and grown in Mix and Go E. coli (Zymo Research, Irvine, CA, USA) in 5 ml Luria-Bertani broth containing selection antibiotic and prepared using the QIAprep Spin Miniprep Kit (Qiagen 27106), per the manufacturer’s instructions.

Techniques: Expressing

Journal: The Journal of Craniofacial Surgery

Article Title: FSTL1 Can Be a Promising Target in TMJ Osteoarthritis via Regulating Chondrocyte Mitophagy and Apoptosis

doi: 10.1097/SCS.0000000000010906

Figure Lengend Snippet: (A) Typical CBCT scan of TMD patients without OA. The red arrow showed the relatively normal bone margins. (B) Typical CBCT scan of TMD patients with OA. The red arrow showed the bone resorption in condyle. (C) The ELISA results of FSTL1 in TMJ auricular synovium fluid. (D) The ROC curve of the diagnosis value FSTL1 concentration in TMJ OA progress. CBCT indicates cone beam computed tomography; FSTL1, follistatin-like protein 1; OA, osteoarthritis; ROC, receiver operating characteristic; TMD, temporomandibular joint disorder; TMJ, temporomandibular joint.

Article Snippet: The used antibodies were as followed, FSTL1 (1:200, Proteintech), ACAN (1:200, Proteintech), COL2A1(1:200, Millipore), and MMP13 (1:200, Abclonal).

Techniques: Enzyme-linked Immunosorbent Assay, Biomarker Discovery, Concentration Assay, Computed Tomography

Journal: The Journal of Craniofacial Surgery

Article Title: FSTL1 Can Be a Promising Target in TMJ Osteoarthritis via Regulating Chondrocyte Mitophagy and Apoptosis

doi: 10.1097/SCS.0000000000010906

Figure Lengend Snippet: (A) The Rt-qPCR results of FSTL1 with the targeting plasmid transfection. (B) The western blot results of FSTL1 with the targeting plasmid transfection. (C) The Rt-qPCR results of the mRNA fold change of related typical markers to chondrocyte synthesis and catabolism as well as under the inflammatory stimulation. (D) The western blot results of changes of related typical markers to chondrocyte synthesis and catabolism as well as under the inflammatory stimulation. (E) The safranine O staining of chondrogenic cells with / without FSTL1 overexpression. FSTL1 indicates follistatin-like protein 1; Rt-qPCR, quantitative reverse transcription polymerase chain reaction.

Article Snippet: The used antibodies were as followed, FSTL1 (1:200, Proteintech), ACAN (1:200, Proteintech), COL2A1(1:200, Millipore), and MMP13 (1:200, Abclonal).

Techniques: Quantitative RT-PCR, Plasmid Preparation, Transfection, Western Blot, Staining, Over Expression, Reverse Transcription, Polymerase Chain Reaction

Journal: The Journal of Craniofacial Surgery

Article Title: FSTL1 Can Be a Promising Target in TMJ Osteoarthritis via Regulating Chondrocyte Mitophagy and Apoptosis

doi: 10.1097/SCS.0000000000010906

Figure Lengend Snippet: (A, B) The mitosox probe staining and the quantified analysis of the mitochondrial morphology and function with / without FSTL1 overexpression. (C, D) The western blot and the quantified analysis results of mitochondrial function with / without FSTL1 overexpression and inflammatory stimulation. (E, F) Flow cytometry analysis of the chondrocytes apoptosis rate with / without FSTL1 overexpression. (G, H) Flow cytometry analysis of the chondrocytes cell cycle with / without FSTL1 overexpression. FSTL1 indicates follistatin-like protein 1.

Article Snippet: The used antibodies were as followed, FSTL1 (1:200, Proteintech), ACAN (1:200, Proteintech), COL2A1(1:200, Millipore), and MMP13 (1:200, Abclonal).

Techniques: Staining, Over Expression, Western Blot, Flow Cytometry

Journal: The Journal of Craniofacial Surgery

Article Title: FSTL1 Can Be a Promising Target in TMJ Osteoarthritis via Regulating Chondrocyte Mitophagy and Apoptosis

doi: 10.1097/SCS.0000000000010906

Figure Lengend Snippet: (A) The Rt-qPCR results mRNA fold change with the targeted si-FSTL1. (B) The western blot results of FSTL1 with the targeting si-RNA and its impact of typical markers to chondrocyte synthesis and catabolism. (C, D) The mitosox probe and IF staining of mt-COⅡ and the quantified analysis of the mitochondrial morphology and function with / without downregulation of FSTL1 under inflammatory stimulation. (E, F) The western blot and the quantified analysis results of mitochondrial function with / without downregulation of FSTL1 and inflammatory stimulation. (G, H) Flow cytometry analysis of the chondrocytes apoptosis rate with / without FSTL1 downregulation. FSTL1 indicates follistatin-like protein 1; Rt-qPCR, quantitative reverse transcription polymerase chain reaction

Article Snippet: The used antibodies were as followed, FSTL1 (1:200, Proteintech), ACAN (1:200, Proteintech), COL2A1(1:200, Millipore), and MMP13 (1:200, Abclonal).

Techniques: Quantitative RT-PCR, Western Blot, Staining, Flow Cytometry, Reverse Transcription, Polymerase Chain Reaction

Journal: The Journal of Craniofacial Surgery

Article Title: FSTL1 Can Be a Promising Target in TMJ Osteoarthritis via Regulating Chondrocyte Mitophagy and Apoptosis

doi: 10.1097/SCS.0000000000010906

Figure Lengend Snippet: (A, B) HE and safranin O/Fast green staining of the cartilage in the sham group and UAC-induced OA rats with si-NC or si-FSTL1 injected group. Scale bar=100 μm. (C) IHC staining of the cartilage in the sham group and UAC-induced OA rats with si-NC or si-FSTL1 injected group. Scale bar=100 μm in the larger field of view and 50 μm in the smaller field of view. (D) The gross OARSI score of the cartilage condition in the sham group and UAC-induced OA rats with si-NC or si-FSTL1 injected group. (E) The statistical analysis of the cartilage IHC staining in the sham group and UAC-induced OA rats with si-NC or si-FSTL1 injected group. FSTL1 indicates follistatin-like protein 1; HE, hematoxylin-eosin; IHC, immunohistochemical; OA, osteoarthritis; UAC, unilateral anterior crossbite.

Article Snippet: The used antibodies were as followed, FSTL1 (1:200, Proteintech), ACAN (1:200, Proteintech), COL2A1(1:200, Millipore), and MMP13 (1:200, Abclonal).

Techniques: Staining, Injection, Immunohistochemistry, Immunohistochemical staining

Journal: European Journal of Nuclear Medicine and Molecular Imaging

Article Title: [ 68 Ga]Ga-NODAGA-RGD post MI reflects activated fibroblasts rather than angiogenesis

doi: 10.1007/s00259-025-07489-4

Figure Lengend Snippet: Violin-plot visualization of gene markers for injury-activated fibroblasts at day 5 post MI (Fstl1, Col1a1, Col3a1, Col5a1) comparing relative gene expression between the experimental groups and cell types. Median values are visualized by a black horizontal bar. ( n = 5 per group)

Article Snippet: Primary antibodies targeting FSTL1 (R&D Systems, AF1738, dilution 1:100), Integrin (Abcam, ab179473, dilution 1:100) CD31 (R&D Systems, AF3628, dilution 1:100) were applied overnight at 4°.

Techniques: Gene Expression

Journal: European Journal of Nuclear Medicine and Molecular Imaging

Article Title: [ 68 Ga]Ga-NODAGA-RGD post MI reflects activated fibroblasts rather than angiogenesis

doi: 10.1007/s00259-025-07489-4

Figure Lengend Snippet: Expression pattern of α v β 3 -integrin, Fstl1 and CD31 in hearts at day 5 after permanent ligation of the LAD in both remote and infarcted myocardium, and the SHAM group. α v β 3 -integrin and Fstl1 are highly expressed within the infarct area ( A ), while profound less expression was detected in the remote area, comparable to the SHAM group ( B , C ). CD31 is expressed within the scar ( D ), but shows basal expression levels in the remote area ( E ) – similar to the SHAM group ( F ). Co-localization of Fstl1 and α v β 3 -integrin was quantified in co-labelled heart sections ( G ). Regions of Interest (ROI) were analyzed according to a positive fluorescence signal for both proteins ( H ). All α v β 3 -integrin positive ROIs were contained in the Fstl1-positive ROI population ( I ). Scale bar: 100 μm

Article Snippet: Primary antibodies targeting FSTL1 (R&D Systems, AF1738, dilution 1:100), Integrin (Abcam, ab179473, dilution 1:100) CD31 (R&D Systems, AF3628, dilution 1:100) were applied overnight at 4°.

Techniques: Expressing, Ligation, Fluorescence

Journal: Life Science Alliance

Article Title: Multi-region proteome analysis quantifies spatial heterogeneity of prostate tissue biomarkers

doi: 10.26508/lsa.201800042

Figure Lengend Snippet: Six proteins (ACPP, ABCF1, NUP93, CUTA, CRAT, and FSTL1) were measured using IHC in a TMA containing tissue samples from 83 patients from an independent cohort, including 35 patients with BPH and 48 patients with prostate ADCA.

Article Snippet: The following primary antibodies were used to stain 4-μm slides of the TMA using the Ventana Benchmark (Roche Ventana Medical Systems, Inc.) automated staining system: ACTR1B (1:400; abcam, 60 min pretreatment), Desmin/DES (1:20; Dako A/S, 16 min pretreatment), KLK3/PSA (1: 10000; Dako A/S) and GDF15 (1:50; Biorbyt, 30 min pretreatment), ACPP (1:2000; DAKO A/S), ABCF1 (1:50; Novus Biologicals, 90 min pretreatment), NUP93 (1:50; Novus Biologicals, 60 min pretreatment), CUTA (1:100; Lifespan Biosciences, 60 min pretreatment), CRAT (1:100; Atlas Antibodies, 30 min pretreatment), and FSTL1 (1:100; Atlas Antibodies, 16 min pretreatment).

Techniques:

Journal: Stroke

Article Title: Follistatin-Like 1 Attenuates Apoptosis via Disco-Interacting Protein 2 Homolog A/Akt Pathway After Middle Cerebral Artery Occlusion in Rats

doi: 10.1161/strokeaha.114.006092

Figure Lengend Snippet: Figure 4. Follistatin-like 1 (FSTL1) decreased apoptosis by increasing Akt phosphorylation 24 hours after middle cerebral artery occlusion (MCAO). A, Immunofluorescence double staining of terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL)–positive cells (green) and neurons (neuronal nuclei [NeuN], red). B, TUNEL-positive neurons are decreased by FSTL1 and increased by FSTL1-siRNA. Neurological score is improved by FSTL1 and reduced by FSTL1-siRNA. C, Western blot analysis of FSTL1 expression. D, Western blot analysis of phosphorylated Akt (p-Akt). Akt phosphorylation is increased by FSTL1 and reversed by FSTL1-siRNA. Disco-interacting pro- tein 2 homolog A (DIP2A; 96 kDa), FSTL1 (35 kDa), and β-actin (42 kDa) were run on the same gel. n=6 for each group in Western blot and immunohistochemistry. &P<0.05 vs MCAO+vehicle, #P<0.05 vs MCAO+scramble-siRNA. Bars, 100 μm.

Article Snippet: To test whether delivery of FSTL1 protein affects acute brain ischemic injury in rats, male Sprague-Dawley rats were injected intracerebroventricularly with either 1 of 2 dosages of recombinant FSTL1 protein (R&D Systems, 100 or 300 mg/kg) or vehicle (0.1 mol/L PBS) at 1 hour after reperfusion. siRNA Injection Three different formats of FSTL1-small interfering RNA (siRNA) (OriGene Technologies) or DIP2A-siRNA (Santa Cruz Biotechnology) were applied 48 hours before MCAO by intracerebroventricular injection (ICV) as previously described.10 A scalp incision was made along the midline, and a burr hole (1 mm) was drilled into the skull above the right hemisphere (1.0 mm lateral of the bregma).

Techniques: Phospho-proteomics, Immunofluorescence, Double Staining, TUNEL Assay, Western Blot, Expressing, Immunohistochemistry

Journal: Stroke

Article Title: Follistatin-Like 1 Attenuates Apoptosis via Disco-Interacting Protein 2 Homolog A/Akt Pathway After Middle Cerebral Artery Occlusion in Rats

doi: 10.1161/strokeaha.114.006092

Figure Lengend Snippet: Figure 6. Follistatin-like 1 (FSTL1) pre- vented apoptosis though the disco- interacting protein 2 homolog A (DIP2A)/ Akt pathway after middle cerebral artery occlusion (MCAO) in rats. Western blots for DIP2A, Akt, and cleaved caspase-3 (CC3; A) showed that DIP2A-siRNA pre- vented the expression of DIP2A after FSTL1 treatment (B), reduced Akt phos- phorylation (C), and increased the expres- sion of CC3 (D). n=6 for each group. &P<0.05 vs MCAO+vehicle; @P<0.05 vs MCAO+FSTL1+scramble-siRNA.

Article Snippet: To test whether delivery of FSTL1 protein affects acute brain ischemic injury in rats, male Sprague-Dawley rats were injected intracerebroventricularly with either 1 of 2 dosages of recombinant FSTL1 protein (R&D Systems, 100 or 300 mg/kg) or vehicle (0.1 mol/L PBS) at 1 hour after reperfusion. siRNA Injection Three different formats of FSTL1-small interfering RNA (siRNA) (OriGene Technologies) or DIP2A-siRNA (Santa Cruz Biotechnology) were applied 48 hours before MCAO by intracerebroventricular injection (ICV) as previously described.10 A scalp incision was made along the midline, and a burr hole (1 mm) was drilled into the skull above the right hemisphere (1.0 mm lateral of the bregma).

Techniques: Western Blot, Expressing

Journal: Stroke

Article Title: Follistatin-Like 1 Attenuates Apoptosis via Disco-Interacting Protein 2 Homolog A/Akt Pathway After Middle Cerebral Artery Occlusion in Rats

doi: 10.1161/strokeaha.114.006092

Figure Lengend Snippet: Figure 5. Follistatin-like 1 (FSTL1) decreased neuron apoptosis after middle cerebral artery occlusion (MCAO) was dependent on its receptor, disco- interacting protein 2 homolog A (DIP2A). DIP2A-siRNA increased neuron apop- tosis (A and B) and increased neuro- logical deficits (C) in FSTL1-treated rats after MCAO. n=6 for each group. &P<0.05 vs MCAO+vehicle; @P<0.05 vs MCAO+FSTL1+scramble-siRNA. Bars, 100 μm.

Article Snippet: To test whether delivery of FSTL1 protein affects acute brain ischemic injury in rats, male Sprague-Dawley rats were injected intracerebroventricularly with either 1 of 2 dosages of recombinant FSTL1 protein (R&D Systems, 100 or 300 mg/kg) or vehicle (0.1 mol/L PBS) at 1 hour after reperfusion. siRNA Injection Three different formats of FSTL1-small interfering RNA (siRNA) (OriGene Technologies) or DIP2A-siRNA (Santa Cruz Biotechnology) were applied 48 hours before MCAO by intracerebroventricular injection (ICV) as previously described.10 A scalp incision was made along the midline, and a burr hole (1 mm) was drilled into the skull above the right hemisphere (1.0 mm lateral of the bregma).

Techniques: