Journal: ACS Applied Nano Materials

Article Title: Antifibrotic and Pro-regenerative Effects of SMAD3 siRNA and Collagen I mRNA-Loaded Lipid Nanoparticles in Human Tenocytes

doi: 10.1021/acsanm.4c02996

Figure Lengend Snippet: Schematic illustration of the one-step microfluidics coflow nanoprecipitation method developed to produce LNPs loaded simultaneously with SMAD3 siRNA and collagen I mRNA. Image created with Biorender.com .

Article Snippet: The probes used in the assay were from Thermo Fisher Scientific and predesigned: 18S (4333760T), human Transforming Growth Factor-b ( TGFB1 , Hs00998133_m1), collagen type III alpha 1 chain ( COL3A1 , Hs00943809_m1), bone morphogenetic protein 2 ( BMP2 , Hs00154192_m1), and SMAD family member 3 (SMAD3 Hs00969210_m1).

Techniques:

Journal: ACS Applied Nano Materials

Article Title: Antifibrotic and Pro-regenerative Effects of SMAD3 siRNA and Collagen I mRNA-Loaded Lipid Nanoparticles in Human Tenocytes

doi: 10.1021/acsanm.4c02996

Figure Lengend Snippet: Physicochemical characterization of the optimized empty LNPs, SMAD3 siRNA LNPs, COL1A1 mRNA LNPs, and dual-loaded LNPs regarding the (A) size (nanometers), (B) PDI, and (C) zeta potential (mV), and EE (D). Colloidal stability of empty LNPs in tenocytes’ cell culture medium and 1× PBS in terms of (E) size and (F) PDI. All the samples were analyzed after dialysis and the results are presented as mean ± s.d. ( n ≥ 3). For statistical analysis, a paired Student’s t -test was used, and p -values were set at probability ** p < 0.01.

Article Snippet: The probes used in the assay were from Thermo Fisher Scientific and predesigned: 18S (4333760T), human Transforming Growth Factor-b ( TGFB1 , Hs00998133_m1), collagen type III alpha 1 chain ( COL3A1 , Hs00943809_m1), bone morphogenetic protein 2 ( BMP2 , Hs00154192_m1), and SMAD family member 3 (SMAD3 Hs00969210_m1).

Techniques: Zeta Potential Analyzer, Cell Culture

Journal: ACS Applied Nano Materials

Article Title: Antifibrotic and Pro-regenerative Effects of SMAD3 siRNA and Collagen I mRNA-Loaded Lipid Nanoparticles in Human Tenocytes

doi: 10.1021/acsanm.4c02996

Figure Lengend Snippet: RT-qPCR study to assess the gene expression of (A) SMAD3 , (B) BMP2 , (C) COL3A1 , and (D) TGFB1 in human tenocytes. Data show the fold increase values compared to the only cells control ± s.d. ( n ≥ 3). For statistical analysis, an ordinary one-way ANOVA followed by a Dunnett posthoc test was used. The significance levels were set at the probabilities of *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001 for comparison with only cells (negative control).

Article Snippet: The probes used in the assay were from Thermo Fisher Scientific and predesigned: 18S (4333760T), human Transforming Growth Factor-b ( TGFB1 , Hs00998133_m1), collagen type III alpha 1 chain ( COL3A1 , Hs00943809_m1), bone morphogenetic protein 2 ( BMP2 , Hs00154192_m1), and SMAD family member 3 (SMAD3 Hs00969210_m1).

Techniques: Quantitative RT-PCR, Gene Expression, Control, Comparison, Negative Control

Journal: ACS Applied Nano Materials

Article Title: Antifibrotic and Pro-regenerative Effects of SMAD3 siRNA and Collagen I mRNA-Loaded Lipid Nanoparticles in Human Tenocytes

doi: 10.1021/acsanm.4c02996

Figure Lengend Snippet: (A) Quantitative analysis of collagen I production using MetaXpress software. The results represent the sum of the intensity values of Texas Red divided by the count of cells. The 32 images were analyzed per well and per sample. (B) Flow cytometry analysis of SMAD3 expression after intracellular staining. For statistical analysis, a one-way ANOVA followed by a Dunnett posthoc test was used in (A) and (B). The significance levels were set at the probabilities of *p < 0.05, and ****p < 0.0001 for comparison with only cells (A, B), mRNA (A), and siRNA (B).

Article Snippet: The probes used in the assay were from Thermo Fisher Scientific and predesigned: 18S (4333760T), human Transforming Growth Factor-b ( TGFB1 , Hs00998133_m1), collagen type III alpha 1 chain ( COL3A1 , Hs00943809_m1), bone morphogenetic protein 2 ( BMP2 , Hs00154192_m1), and SMAD family member 3 (SMAD3 Hs00969210_m1).

Techniques: Software, Flow Cytometry, Expressing, Staining, Comparison

Journal: Cell stem cell

Article Title: A hierarchy of proliferative and migratory keratinocytes maintains the tympanic membrane

doi: 10.1016/j.stem.2020.10.006

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit monoclonal anti-α-Smooth Muscle Actin (Acta2); 1:100 , Cell Signaling Technology , Cat# 19245, RRID: 2734735.

Techniques: Membrane, Drug discovery, RNAscope, Multiplex Assay, Imaging, Software

Journal: Burns & Trauma

Article Title: Overexpression of miR-101 suppresses collagen synthesis by targeting EZH2 in hypertrophic scar fibroblasts

doi: 10.1093/burnst/tkab038

Figure Lengend Snippet: Overexpression of miR-101 decreased fibrosis-related protein expression and proliferation. miR-101 RNA levels were analyzed using qRT-PCR normalized to U6 in HS ( a ) and HSF ( b ), compared with NS and NSF. Each data point was normalized against its corresponding U6 level. ( c ) FISH staining for miR-101 using serial sections of clinical HS and paired NS. Scale bar = 50 μm. ( d ) qRT-PCR analysis of the effects of mimics control, miR-101 mimics, inhibitor control and miR-101 inhibitor on the level of miR-101. qRT-PCR analysis ( e , f ) and representative immunoblots ( g ) showing the mRNA level and protein level changes of Col1, Col3 and α-SMA in HSF transfected with mimics control, miR-101 mimics, inhibitor control and miR-101 inhibitor. ( h , i ) Col1 and Col3 supernatant protein levels were determined using ELISA in the same treatment. Data are expressed as the mean ± SD of three independent experiments. * p < 0.05, * * p < 0.01. qRT-PCR quantitative reverse-transcriptase polymerase chain reaction, U6 U6 small nuclear 1, HS hypertrophic scar, HSF hypertrophic scar fibroblast, NS normal skin, NSF normal skin fibroblast, FISH fluorescence in situ hybridization, ELISA enzyme-linked immunosorbent assay.

Article Snippet: Antibodies were as follows: anti-Col1 (rabbit, 1:1000; Abcam), anti-Col3 (rabbit, 1:3000; Abcam), anti-α-SMA (rabbit, 1:1000; Abcam), anti-EZH2 (rabbit, 1:1000; Abcam) and anti-GAPDH (Rabbit, 1:3000; Abcam).

Techniques: Over Expression, Expressing, Quantitative RT-PCR, Staining, Western Blot, Transfection, Enzyme-linked Immunosorbent Assay, Polymerase Chain Reaction, Fluorescence, In Situ Hybridization

Journal: Burns & Trauma

Article Title: Overexpression of miR-101 suppresses collagen synthesis by targeting EZH2 in hypertrophic scar fibroblasts

doi: 10.1093/burnst/tkab038

Figure Lengend Snippet: The role of EZH2 in regulating collagen and α-SMA expression in HSF. EZH2 mRNA ( a ) and protein expression ( b ) were measured using qRT-PCR and western blot analyses, respectively, in siRNA-EZH2 and siRNA-control transfected HSF. ( c ) RNA sequencing profiling of HSF in which EZH2 was knocked down. Shades of red represent increased gene expression; shades of blue represent decreased gene expression. ( d ) Pathway enrichment was done using the ClueGO and CluePedia plugins of the Cytoscape software. This analysis identified seven enriched pathways based on statistical analysis, with no duplication between clusters. Cluster one mainly includes positive regulation of collagen production ( e ) and fibroblasts responsible for TGF-β activation ( f ). Immunoblotting ( g ) or qRT-PCR ( h ) assessing the effect of EZH2-siRNA on the expression of Col1, Col3 and α-SMA in HSF. ( i , j ) ELISA of Col1 and Col3 supernatant protein levels in EZH2 knocked down HSF. EZH2 enhancer of zeste homolog 2, qRT-PCR quantitative reverse-transcriptase polymerase chain reaction, HSF hypertrophic scar fibroblast, ELISA enzyme-linked immunosorbent assay

Article Snippet: Antibodies were as follows: anti-Col1 (rabbit, 1:1000; Abcam), anti-Col3 (rabbit, 1:3000; Abcam), anti-α-SMA (rabbit, 1:1000; Abcam), anti-EZH2 (rabbit, 1:1000; Abcam) and anti-GAPDH (Rabbit, 1:3000; Abcam).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Transfection, RNA Sequencing Assay, Software, Activation Assay, Enzyme-linked Immunosorbent Assay, Polymerase Chain Reaction

Journal: Burns & Trauma

Article Title: Overexpression of miR-101 suppresses collagen synthesis by targeting EZH2 in hypertrophic scar fibroblasts

doi: 10.1093/burnst/tkab038

Figure Lengend Snippet: miR-101 targets EZH2 in HSF. Expression levels of miR-101 are negatively correlated with EZH2 expression in HS ( a ) and HSF ( b ). Pearson’s correlation analysis of the relative expression levels of miR-101 (normalized to U6) and EZH2 (normalized to GAPDH) was determined using qRT-PCR in nine paired HS and HSF samples. ( c ) Binding of miR-101 to the wild type EZH2 3′ UTR (WT-EZH2-UTR) and the designed mutant (Mut-EZH2-UTR). ( d ) Dual-luciferase reporter assay showing that miR-101 mimics reduced luciferase activity in HEK293A cells after transfection with WT- EZH2-UTR, but not with the Mut- EZH2-UTR vector. Analysis of expression levels of EZH2 in HSF infection with miR-101 and miR-101 inhibitor using qRT-PCR ( e ) and western blot ( f ) analyses. Western blot analysis of Col1, Col3 and α-SMA protein expression in HSF treated with miR-101 mimics or infected with lentivirus overexpressing EZH2. ( h ) Western blot analysis of Col1, Col3 and α-SMA protein expression in HSF treated with miR-101 inhibitor or EZH2 siRNA. ELISA analysis of Col1 ( i ) and Col3 ( j ) supernatant protein level treated with miR-101 mimics or infected with lentivirus overexpressing EZH2. ELISA analysis of Col1 ( k ) and Col3 ( l ) supernatant protein level treated with miR-101 inhibitor or EZH2 siRNA. EZH2 enhancer of zeste homolog 2, HS hypertrophic scar, HSF hypertrophic scar fibroblast, U6 U6 small nuclear 1, UTR untraslated region, WT wild type, Mut mutant, HEK293A human embryonic kidney 293A cell, ELISA enzyme-linked immunosorbent assay

Article Snippet: Antibodies were as follows: anti-Col1 (rabbit, 1:1000; Abcam), anti-Col3 (rabbit, 1:3000; Abcam), anti-α-SMA (rabbit, 1:1000; Abcam), anti-EZH2 (rabbit, 1:1000; Abcam) and anti-GAPDH (Rabbit, 1:3000; Abcam).

Techniques: Expressing, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Reporter Assay, Activity Assay, Transfection, Plasmid Preparation, Infection, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Asiaticoside Combined With Carbon Ion Implantation to Improve the Biocompatibility of Silicone Rubber and to Reduce the Risk of Capsule Contracture

doi: 10.3389/fbioe.2022.810244

Figure Lengend Snippet: Effect of asiaticoside on TGF-β1-induced transformation of fibroblasts into myofibroblasts and collagen deposition. Immunofluorescence staining of human fibroblasts on silicone rubber (SR) and carbon silicone rubber (C-SR) with Col-1A1 and α-SMA (100X).

Article Snippet: The primary antibodies were as follows: Smad2/3 (1:500, Affinity, United States), vimentin (1:500, Proteintech, China), α-SMA (1:500, Proteintech, China), Col-1A1 (1:500, Novus, United States), TGF-β1 (1:500, Affinity, United States), TβRI (1:500, Abcam, United States), TβRII (1:500, Abcam, United States), Smad2/3 (1:500, Affinity, United States), p-Smad2/3 (1:500, Affinity, United States), and β-actin (1:1,000, Abcam, United States).

Techniques: Transformation Assay, Immunofluorescence, Staining

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Asiaticoside Combined With Carbon Ion Implantation to Improve the Biocompatibility of Silicone Rubber and to Reduce the Risk of Capsule Contracture

doi: 10.3389/fbioe.2022.810244

Figure Lengend Snippet: Effect of asiaticoside on transformation of fibroblasts into myofibroblasts, collagen deposition and changes in the TGF-β/Smad signaling pathway in vivo . (A) Representative Western blot analysis of TGF-β1, TβRI, TβRII, Col-1A1, Smad2/3, p-Smad2/3 and α-SMA. One representative blot of three experiments is presented. β-actin was used as loading control. (B–G) Protein signals in A were determined by densitometric analysis of Western blotting using ImageJ software (NIH). Data are reported as the mean ± standard error ( n = 3, ∗ p < 0.05).

Article Snippet: The primary antibodies were as follows: Smad2/3 (1:500, Affinity, United States), vimentin (1:500, Proteintech, China), α-SMA (1:500, Proteintech, China), Col-1A1 (1:500, Novus, United States), TGF-β1 (1:500, Affinity, United States), TβRI (1:500, Abcam, United States), TβRII (1:500, Abcam, United States), Smad2/3 (1:500, Affinity, United States), p-Smad2/3 (1:500, Affinity, United States), and β-actin (1:1,000, Abcam, United States).

Techniques: Transformation Assay, In Vivo, Western Blot, Control, Software

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Asiaticoside Combined With Carbon Ion Implantation to Improve the Biocompatibility of Silicone Rubber and to Reduce the Risk of Capsule Contracture

doi: 10.3389/fbioe.2022.810244

Figure Lengend Snippet: Effect of asiaticoside on the transformation of fibroblasts into myofibroblasts, collagen deposition and fibroblast proliferation in vivo . Immunohistochemical staining of tissue around silicone rubber (SR) and carbon silicone rubber (C-SR) with α-SMA, vimentin, PCNA and COL-1A1 (100X).

Article Snippet: The primary antibodies were as follows: Smad2/3 (1:500, Affinity, United States), vimentin (1:500, Proteintech, China), α-SMA (1:500, Proteintech, China), Col-1A1 (1:500, Novus, United States), TGF-β1 (1:500, Affinity, United States), TβRI (1:500, Abcam, United States), TβRII (1:500, Abcam, United States), Smad2/3 (1:500, Affinity, United States), p-Smad2/3 (1:500, Affinity, United States), and β-actin (1:1,000, Abcam, United States).

Techniques: Transformation Assay, In Vivo, Immunohistochemical staining, Staining

Journal: Stem Cell Research & Therapy

Article Title: Microparticles derived from human erythropoietin mRNA-transfected mesenchymal stem cells inhibit epithelial-to-mesenchymal transition and ameliorate renal interstitial fibrosis

doi: 10.1186/s13287-020-01932-z

Figure Lengend Snippet: qRT-PCR primers

Article Snippet: After blocking with 5% skim milk, the membranes were incubated with primary antibodies against E-cadherin (1:500; BD Biosciences, Bedford, UK), α-SMA (1:10000; R&D, Minneapolis, IL, USA), fibronectin (1:1000; Santa Cruz, CA, USA), phospho-p38 (1:1000; Santa Cruz), p38 (1:1000; Abcam, Cambridge, UK), phospho-Smad2 (1:500; Millipore), Smad2 (1:1000; Santa Cruz), phospho-Smad3 (1:500; Abcam), Smad3 (1:1000; Cell Signaling Technology, Danvers, MA, USA), and GAPDH (1:1000; Cell Signaling Technology, Danvers, MA, USA).

Techniques:

Journal: Stem Cell Research & Therapy

Article Title: Microparticles derived from human erythropoietin mRNA-transfected mesenchymal stem cells inhibit epithelial-to-mesenchymal transition and ameliorate renal interstitial fibrosis

doi: 10.1186/s13287-020-01932-z

Figure Lengend Snippet: MPs and rhEPO attenuate TGF-β1-induced EMT in MDCK cells. A Morphological changes in TGF-β1-treated MDCK cells observed by phase contrast microscopy. Compared to control MDCK cells (a), TGF-β1 treatment changed MDCK cell morphology from cuboidal to an elongated spindle-like shape (b). These morphological changes were reversed by co-treatment with MOCK-MPs (c), hEPO-MPs (d), and rhEPO (e). Fluorescence microscopy of red fluorescent CellTracker™-labeled MP incorporation into MDCK cells (m, n, r, s). Blue staining represents nuclear counterstaining with DAPI (f–j, p–t). B RT-PCR of hEPO mRNA expression in MDCK cells co-treated with hEPO-MPs, demonstrating that MPs mediated the horizontal transfer of hEPO mRNA into target MDCK cells. C Immunofluorescence microscopy of E-cadherin, vimentin, α-SMA, and fibronectin in MDCK cells after co-treatment with TGF-β1 and MPs or rhEPO, indicating reduced E-cadherin immunostaining intensity in response to TGF-β1-treatment (b) and its reversal by co-treatment with MOCK-MPs (c), hEPO-MPs (d), and rhEPO (e). TGF-β1 treatment increased vimentin (g), α-SMA (l), and fibronectin (q) immunostaining intensity which was reversed by co-treatment with MOCK-MPs (h, m, r), hEPO-MPs (i, n, s), and rhEPO (j, o, t). Magnification, × 400. Scale bars, 100 μm (white)

Article Snippet: After blocking with 5% skim milk, the membranes were incubated with primary antibodies against E-cadherin (1:500; BD Biosciences, Bedford, UK), α-SMA (1:10000; R&D, Minneapolis, IL, USA), fibronectin (1:1000; Santa Cruz, CA, USA), phospho-p38 (1:1000; Santa Cruz), p38 (1:1000; Abcam, Cambridge, UK), phospho-Smad2 (1:500; Millipore), Smad2 (1:1000; Santa Cruz), phospho-Smad3 (1:500; Abcam), Smad3 (1:1000; Cell Signaling Technology, Danvers, MA, USA), and GAPDH (1:1000; Cell Signaling Technology, Danvers, MA, USA).

Techniques: Microscopy, Fluorescence, Labeling, Staining, Reverse Transcription Polymerase Chain Reaction, Expressing, Immunofluorescence, Immunostaining

Journal: Stem Cell Research & Therapy

Article Title: Microparticles derived from human erythropoietin mRNA-transfected mesenchymal stem cells inhibit epithelial-to-mesenchymal transition and ameliorate renal interstitial fibrosis

doi: 10.1186/s13287-020-01932-z

Figure Lengend Snippet: Effect of MPs or rhEPO on EMT markers in MDCK cells. A – C qRT-PCR of E-cadherin, vimentin, and α-SMA mRNA in MDCK cells co-treated with TGF-β1 and MPs or rhEPO. Compared to vehicle-treated MDCK cells, TGF-β1 treatment significantly decreased E-cadherin and increased vimentin and α-SMA mRNA expression. MOCK-MPs, hEPO-MPs, and rhEPO reversed these changes in E-cadherin, vimentin, and α-SMA mRNA expression. D Western blotting for E-cadherin, α-SMA, and fibronectin protein levels in MDCK cells co-treated with TGF-β1 and MPs or rhEPO. Compared to vehicle-treated MDCK cells, TGF-β1 significantly decreased E-cadherin expression ( E ) and increased α-SMA and fibronectin expression ( F , G ). Co-treatment with MOCK-MPs, hEPO-MPs, and rhEPO significantly ameliorated these changes induced by TGF-β1. Western blotting data were normalized to GAPDH. Molecular weights are shown in kDa. Data are presented as mean ± SD; n = 4~5 for each experimental group. a P < 0.05 vs. vehicle, b P < 0.05 vs. TGF-β1, c P < 0.05 vs. TGF-β1+MOCK-MPs, d P < 0.05 vs. TGF-β1+hEPO-MPs, e P < 0.05 vs. TGF-β1+rhEPO

Article Snippet: After blocking with 5% skim milk, the membranes were incubated with primary antibodies against E-cadherin (1:500; BD Biosciences, Bedford, UK), α-SMA (1:10000; R&D, Minneapolis, IL, USA), fibronectin (1:1000; Santa Cruz, CA, USA), phospho-p38 (1:1000; Santa Cruz), p38 (1:1000; Abcam, Cambridge, UK), phospho-Smad2 (1:500; Millipore), Smad2 (1:1000; Santa Cruz), phospho-Smad3 (1:500; Abcam), Smad3 (1:1000; Cell Signaling Technology, Danvers, MA, USA), and GAPDH (1:1000; Cell Signaling Technology, Danvers, MA, USA).

Techniques: Quantitative RT-PCR, Expressing, Western Blot

Journal: Stem Cell Research & Therapy

Article Title: Microparticles derived from human erythropoietin mRNA-transfected mesenchymal stem cells inhibit epithelial-to-mesenchymal transition and ameliorate renal interstitial fibrosis

doi: 10.1186/s13287-020-01932-z

Figure Lengend Snippet: Anti-fibrotic effects of MPs or rhEPO in UUO kidneys. A Microscopic analysis of Masson’s trichrome and Sirius red-stained UUO kidney sections. Sham-operated control (a, f), obstructed kidney treated with vehicle control (b, g), and UUO kidneys treated with MOCK-MPs (c, h), hEPO-MPs (d, i), and rhEPO (e, j). MPs and rhEPO significantly attenuated renal interstitial fibrosis as quantified using Image J and MetaMorph software ( B , C ). Immunohistochemical analysis of α-SMA expression in sham-operated control (k) and UUO kidneys treated with the vehicle control (l), MOCK-MPs (m), hEPO-MPs (n), and rhEPO (o). MPs and rhEPO significantly attenuated α-SMA expression in obstructed kidneys ( D ). Immunohistochemical analysis of F4/80-positive cells in sham-operated control (p) and UUO kidneys treated with vehicle control (q), MOCK-MPs (r), hEPO-MPs (s), and rhEPO (t). MPs and rhEPO significantly attenuated the infiltration of F4/80-positive macrophage/inflammatory cells in obstructed kidneys. Quantitative assessment of renal F4/80-positive macrophage/inflammatory cells ( E ). Magnification, × 400 and × 1000 (for inserts). Scale bars, 50 μm (black). Data are presented as the mean ± SD; n = 5 for each experimental group. a P < 0.05 vs. sham control, b P < 0.05 vs. UUO+vehicle, c P < 0.05 vs. UUO+MOCK-MPs, d P < 0.05 vs. UUO+hEPO-MPs

Article Snippet: After blocking with 5% skim milk, the membranes were incubated with primary antibodies against E-cadherin (1:500; BD Biosciences, Bedford, UK), α-SMA (1:10000; R&D, Minneapolis, IL, USA), fibronectin (1:1000; Santa Cruz, CA, USA), phospho-p38 (1:1000; Santa Cruz), p38 (1:1000; Abcam, Cambridge, UK), phospho-Smad2 (1:500; Millipore), Smad2 (1:1000; Santa Cruz), phospho-Smad3 (1:500; Abcam), Smad3 (1:1000; Cell Signaling Technology, Danvers, MA, USA), and GAPDH (1:1000; Cell Signaling Technology, Danvers, MA, USA).

Techniques: Staining, Software, Immunohistochemical staining, Expressing

Journal: Stem Cell Research & Therapy

Article Title: Microparticles derived from human erythropoietin mRNA-transfected mesenchymal stem cells inhibit epithelial-to-mesenchymal transition and ameliorate renal interstitial fibrosis

doi: 10.1186/s13287-020-01932-z

Figure Lengend Snippet: Western blotting of α-SMA, collagen I, fibronectin, and TGF-β1 in UUO kidneys. Compared to the sham-operated control, the obstructed kidneys of vehicle-treated mice showed significantly lower E-cadherin ( B ) and higher α-SMA ( C ), collagen I ( D ), fibronectin ( E ), and TGF-β1 ( F ) expression. Treatment with MOCK-MPs, hEPO-MPs, and rhEPO significantly reversed these changes. Protein levels were calculated using NIH Image J software. Western blot data were normalized to GAPDH. Molecular weights are shown in kDa. Data are presented as the mean ± SD; n = 5 for each experimental /group. a P < 0.05 vs. sham control, b P < 0.05 vs. UUO+vehicle, c P < 0.05 vs. UUO+MOCK-MPs, d P < 0.05 vs. UUO+hEPO-MPs, e P < 0.05 vs. UUO+rhEPO

Article Snippet: After blocking with 5% skim milk, the membranes were incubated with primary antibodies against E-cadherin (1:500; BD Biosciences, Bedford, UK), α-SMA (1:10000; R&D, Minneapolis, IL, USA), fibronectin (1:1000; Santa Cruz, CA, USA), phospho-p38 (1:1000; Santa Cruz), p38 (1:1000; Abcam, Cambridge, UK), phospho-Smad2 (1:500; Millipore), Smad2 (1:1000; Santa Cruz), phospho-Smad3 (1:500; Abcam), Smad3 (1:1000; Cell Signaling Technology, Danvers, MA, USA), and GAPDH (1:1000; Cell Signaling Technology, Danvers, MA, USA).

Techniques: Western Blot, Expressing, Software

Journal: Dose-response : a publication of International Hormesis Society

Article Title: Radiation Protection of Polydatin Against Radon Exposure Injury of Epithelial Cells and Mice.

doi: 10.1177/15593258231172271

Figure Lengend Snippet: Figure 4. PD can regulate the expression of EMT markers in radon-exposed cells. (A) The protein expression of E-cad, Vimentin, N-cad, FN1, α-SMA, and Snail in radon exposure model of 16HBE and BEAS-2B cells with directly adding PD. (C) The protein expression of E-cad, Vimentin, N-cad, FN1, α-SMA, and Snail in radon-exposed 16HBE and BEAS-2B cells with PD added before and after radon exposure. (B, D) The expression level of EMT markers in different groups was quantified by Image J software. *: Compared with Rn6, P < .05; * *: Compared with Rn6, P < .01.

Article Snippet: The membrane was sealed with 5% skim milk powder at room temperature and incubated overnight at 4°C with primary antibodies (E-cad (1: 3000, Abcam, USA), Vimentin (1: 5000, Abcam), FN1 (1: 5000, Abcam), Snail (1: 1000, CST, USA), α-SMA (1: 1000, CST), N-cad (1:1000, CST), p-PI3K (1: 500, CST), p-AKT (1: 1000, CST), p-mTOR (1: 800, CST), and GAPDH/ β-tubulin).

Techniques: Expressing, Software

Journal: iScience

Article Title: Profibrotic role of transcription factor SP1 in cross-talk between fibroblasts and M2 macrophages

doi: 10.1016/j.isci.2023.108484

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-Smad7 , Proteintech , cat# 25840-1-AP; RRID: AB_2848137.

Techniques: Recombinant, cDNA Synthesis, CCK-8 Assay, Luciferase, Reporter Gene Assay, Methylation, Mass Spectrometry, Real-time Polymerase Chain Reaction, Knockdown, Plasmid Preparation, Software

Journal: Cell Reports Methods

Article Title: Modeling alcohol-associated liver disease in humans using adipose stromal or stem cell-derived organoids

doi: 10.1016/j.crmeth.2024.100778

Figure Lengend Snippet:

Article Snippet: Smooth Muscle Actin Polyclonal antibody , Proteintech , Cat# 14395-1-AP; RRID: AB_2223009.

Techniques: Control, Virus, Recombinant, Staining, Enzyme-linked Immunosorbent Assay, Diagnostic Assay, Detection Assay, Quantitative RT-PCR, Software

Journal: Journal of the American Society of Nephrology

Article Title: Thrombospondin 2 Functions as an Endogenous Regulator of Angiogenesis and Inflammation in Experimental Glomerulonephritis in Mice

doi: 10.1681/asn.2006080873

Figure Lengend Snippet: Figure 2. During anti-GBM nephritis, the lack of TSP2 neither leads to compensatory upregulation of TSP1 nor influences TGF- activation. During anti-GBM nephritis, TSP1 was de- tected predominantly in cells of Bowman’s capsule, in the periglomerular tubulointerstitium (A), and in some cases within the glomerulus (B), as assessed by immunohistochem- istry (brown staining). Semiquantitative evaluation of cortical TSP1 revealed peak expression at approximately days 14 to 21 in both groups (C). Phosphorylated Smad 2/3, as an indirect indicator of activated TGF-, was evaluated by immunohisto- chemistry followed by computer-assisted analysis using Meta- Vue software (D). Magnifications: 400 in A and B.

Article Snippet: To perform immunostaining, we incubated tissue sections with the following primary and secondary antibodies: 19A2, a murine IgG mAb against proliferating cell nuclear antigen (PCNA; Chemicon, Temecula, CA), an indicator of actively proliferating cells; a rat monoclonal IgG2a to mouse CD4 antigen (Caltag Laboratory, Burlingame, CA); a rat monoclonal IgG2a to mouse CD8a antigen (Caltag Laboratory [18]); F4/80, a murine IgG1 mAb to a surface receptor that is present on monocytes, macrophages, and dendritic cells (Caltag Laboratory); MECA-32, a murine IgG1 mAb that is specific for detecting endothelial cells (a gift from R. Hallmann, University of Münster, Münster, Germany); a murine IgG2 mAb to -smooth muscle actin ( -SMA; Dako, Hamburg, Germany) (21); a rabbit polyclonal antibody to collagen I (Biogenesis, New Fields, UK); a biotinylated polyclonal antibody to human collagen IV (Southern Biotechnology Associates, Birmingham, UK); a murine IgG1 mAb against TSP1 (Dunn, Labortechnik, Asbach, Germany [21]); a rabbit polyclonal antibody to TSP2 (23); and a rabbit polyclonal antibody to phosphorylated Smad 2/3 (Santa Crutz Biotechnology, Santa Cruz, CA [10]).

Techniques: Activation Assay, Staining, Expressing, Immunohistochemistry, Software