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Thermo Fisher beta actin
Beta Actin, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rabbit+anti+%CE%B2+actin/Actin%2C+from+Rabbit+Muscle/pm42176149-52-37-44
Average 95 stars, based on 1 article reviews

beta actin - by Bioz Stars, 2026-09

95/100 stars

Images

Incubation:

Article Title: Downregulation of ROS1 enhances the therapeutic efficacy of arsenic trioxide in acute myeloid leukemia cell lines
Article Snippet: The protein extracts were subjected to 7.5% SDS-PAGE at 200 V for 40 min and the resolved proteins were electro-transferred for 12 min at 18 V to nitrocellulose membranes (Thermo Fisher Scientific, Inc.) using a western blotting semi-dry transfer unit (Hoefer, San Francisco, CA, USA). .. The membranes were blocked for 1 h at room temperature with TBST (10X buffer; 200 mM Tris base, 1.5 M NaCl in MiliQ water; 0.1% Tween-20 was added and pH was adjusted to 7.5 for 1X buffer) containing 5% BSA The membranes were then incubated with one of the primary antibodies (1:200) [rabbit anti-ROS 1 (cat no. MA5-26180; Invitrogen; Thermo Fisher Scientific, Inc.), rabbit anti-Axin (cat no. 34-5900; Invitrogen; Thermo Fisher Scientific, Inc.), rabbit anti-GSK-3β (cat no. A170132; Invitrogen; Thermo Fisher Scientific, Inc.), rabbit anti-APC (cat no. 17-9956-42; Invitrogen; Thermo Fisher Scientific, Inc.), rabbit anti-β-catenin (cat no. 71-2700; Invitrogen; Thermo Fisher Scientific, Inc.), rabbit anti-c-Myc (cat no. MA1-980; Invitrogen; Thermo Fisher Scientific, Inc.), rabbit anti-Cyclin-D1 (cat no. PA5-16777; Invitrogen; Thermo Fisher Scientific, Inc.), rabbit anti-PPAR-α (cat no. PA1-820; Invitrogen; Thermo Fisher Scientific, Inc.), rabbit anti-MMP-7 (cat no. PA1-9069; Invitrogen; Thermo Fisher Scientific, Inc.), rabbit anti-GAPDH (cat no. PA1-988; Invitrogen; Thermo Fisher Scientific, Inc.) and rabbit anti-β-actin (cat no. PA1-183; Invitrogen; Thermo Fisher Scientific, Inc.)] in TBST with 5% BSA overnight at 4°C with gentle shaking. .. Membranes were washed twice with TBST for 10 min each and additionally incubated with secondary antibody (goat anti-rabbit; cat no. {"type":"entrez-nucleotide","attrs":{"text":"A11008","term_id":"492390","term_text":"A11008"}} A11008 ; 1:5,000; Invitrogen; Thermo Fisher Scientific, Inc.) for 1 h at room temperature.

Gentle:

Expressing:

Article Title: G protein-coupled KISS1 receptor is overexpressed in triple negative breast cancer and promotes drug resistance.
Article Snippet: .. Protein expression was examined using antibodies raised against human proteins: rabbit anti-KISS1R (1:4000, Abcam), rabbit anti-KISS1 (1:500; Abcam), rat anti-BCRP (1:150, Abcam), rabbit anti-cleaved-PARP (1: 1000, Cell Signaling), rabbit anti-PARP (1: 1000, Cell Signaling), mouse anti-Histone H3 (1:1000; Millipore); mouse anti-HSP90 (1:1000, Cell Signaling); mouse anti-phosphoptyrosine (PY-20, 1:1000, Millipore); rabbit anti-AXL (1:2000, Cell Signaling), rabbit anti-snail/slug (1:500, Abcam), mouse anti-N-cadherin (1:500, BD Biosciences), rabbit anti-ERK (1:1000, Cell Signaling), rabbit anti-phoshpo-ERK1/2 (1:2000, Cell Signaling), rabbit anti-AKT (1:1000, Cell Signaling), rabbit anti-phospho-AKT (1:1000, Cell Signaling), rabbit anti-survivin (1:1000, ThermoScientific), rabbit anti-SP-1 (1:1000, Millipore), mouse anti-tubulin (1:5000, Abcam) or mouse anti-β -actin (1:1000, ThermoScientific), rabbit anti-β -actin (1:2000, ThermoScientific), GADPH (1:2000, Abcam) and visualized by chemiluminescence. β -actin, GADPH or tubulin expression was used as a loading control. ..

Control:

other:

Article Title: Global characterization of macrophage polarization mechanisms and identification of M2-type polarization inhibitors
Article Snippet: Rabbit anti-β-actin , Lab Vision , Rb-9421-P1, RRID:AB_720056.

Image Search Results

TAPC interacts with VEGFR2 and modulates downstream signaling. (a) Cell viability assay of MC38 cells treated with increasing concentrations of TAPC. (b) Immunoblot analysis of VEGFR2 and key regulators of the PI3K–AKT signaling pathway (PI3K, AKT, and STAT3) in MC38 cells treated with PEG-PO or TAPC (5 and 10 μM). β-Actin was used as a loading control. (c) Pull-down assay of VEGFR2 from MC38 cell lysates using biotinylated TAPC, beads-only sample served as control. (d) Confocal IF imaging of MC38 cells incubated with Cy5.5-labeled TAPC and stained for VEGFR2, nuclei counterstained with DAPI. Scale bars: 20 μm. (e) BLI analysis of TAPC binding to recombinant VEGFR2 using serial concentrations (100, 66.7, 44.4, 29.6, 19.8, 13.2, and 8.8 μM). (f) Molecular dynamics simulations showing predicted protein–ligand complexes (top) and binding pocket visualizations (bottom) of VEGFR2 with TAPC, NDMPFI, MBAMF, and TPFE. (g) Binding free energy calculations of these complexes, including van der Waals, electrostatic, solvation, and total energy components. (h) Extracellular acidification rate (ECAR) of MC38 cells treated with control (0 μM), TAPC (2.5 μM), or TAPC (10 μM), with sequential addition of glucose, oligomycin, and 2-deoxyglucose (2-DG). (i) Quantification of glycolysis and glycolytic capacity in MC38 cells treated with control (0 μM), TAPC (2.5 μM), or TAPC (10 μM) (n = 8). Data are presented as mean ± SEM. Statistical significance was assessed using one-way ANOVA with Tukey's multiple comparisons test; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Journal: Bioactive Materials

Article Title: Aminated fullerene-based nanoplatform enables synergistic VEGFR2-targeted anti-angiogenesis and tumor immunotherapy

doi: 10.1016/j.bioactmat.2026.03.016

Figure Lengend Snippet: TAPC interacts with VEGFR2 and modulates downstream signaling. (a) Cell viability assay of MC38 cells treated with increasing concentrations of TAPC. (b) Immunoblot analysis of VEGFR2 and key regulators of the PI3K–AKT signaling pathway (PI3K, AKT, and STAT3) in MC38 cells treated with PEG-PO or TAPC (5 and 10 μM). β-Actin was used as a loading control. (c) Pull-down assay of VEGFR2 from MC38 cell lysates using biotinylated TAPC, beads-only sample served as control. (d) Confocal IF imaging of MC38 cells incubated with Cy5.5-labeled TAPC and stained for VEGFR2, nuclei counterstained with DAPI. Scale bars: 20 μm. (e) BLI analysis of TAPC binding to recombinant VEGFR2 using serial concentrations (100, 66.7, 44.4, 29.6, 19.8, 13.2, and 8.8 μM). (f) Molecular dynamics simulations showing predicted protein–ligand complexes (top) and binding pocket visualizations (bottom) of VEGFR2 with TAPC, NDMPFI, MBAMF, and TPFE. (g) Binding free energy calculations of these complexes, including van der Waals, electrostatic, solvation, and total energy components. (h) Extracellular acidification rate (ECAR) of MC38 cells treated with control (0 μM), TAPC (2.5 μM), or TAPC (10 μM), with sequential addition of glucose, oligomycin, and 2-deoxyglucose (2-DG). (i) Quantification of glycolysis and glycolytic capacity in MC38 cells treated with control (0 μM), TAPC (2.5 μM), or TAPC (10 μM) (n = 8). Data are presented as mean ± SEM. Statistical significance was assessed using one-way ANOVA with Tukey's multiple comparisons test; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Article Snippet: Antibodies were listed as follows: Anti-VEGF Receptor 2 antibody [EPRER16Y] (Abcam, Cat: ab134191), Anti-PI 3 Kinase catalytic subunit gamma (Abcam, Cat: ab302958), Anti-AKT (phosphor T308) antibody (Abcam, Cat: ab38449), Anti-STAT3 antibody [EPR787Y] (Abcam, Cat: ab68153), β-Actin (13E5) rabbit mAb (CST, Cat: #4970), Anti-CD31 antibody [EPR17260-263] (Abcam, Cat: ab222783), FITC anti-mouse CD45 (Biolegend, Cat: 103108), PerCP/Cyanine5.5 anti-mouse CD4 (Biolegend, Cat: 100434), FOXP3 Monoclonal Antibody (NRRF-30), PE, eBioscience (Thermo, Cat: 12-4771-82), CD3 (Abcam, Cat: ab16669), CD4 (Servicebio, Cat: GB15064).

Techniques: Viability Assay, Western Blot, Control, Pull Down Assay, Imaging, Incubation, Labeling, Staining, Binding Assay, Recombinant

In vivo anti-tumor and anti-angiogenic effects of TAPC@CNPs. (a) Schematic illustration of the therapeutic study in Balb/c mice bearing subcutaneous MC38 tumors (n = 7). (b) Body weights of mice during treatment. (c) Photographs of excised tumors collected at endpoint. (d) Tumor growth curves during treatment. Tumor volume was calculated using the formula (length × width 2 )/2. (e) Tumor weights measured at endpoint. (f) Immunoblot analysis of VEGFR2 expression in tumor lysates from different treatment groups, β-actin was used as a reference protein. (g) IHC staining of CD31 in tumor sections from different treatment groups. Scale bar, 100 μm. (h) H&E staining of major organs (heart, liver, spleen, lung, kidney) and tumor tissues. (i) Serum ALT and AST levels measured at endpoint. Data are presented as mean ± SEM. Statistical analysis was performed by one-way ANOVA with Tukey's multiple comparisons test, ns indicates not significant, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗∗p < 0.0001.

Journal: Bioactive Materials

Article Title: Aminated fullerene-based nanoplatform enables synergistic VEGFR2-targeted anti-angiogenesis and tumor immunotherapy

doi: 10.1016/j.bioactmat.2026.03.016

Figure Lengend Snippet: In vivo anti-tumor and anti-angiogenic effects of TAPC@CNPs. (a) Schematic illustration of the therapeutic study in Balb/c mice bearing subcutaneous MC38 tumors (n = 7). (b) Body weights of mice during treatment. (c) Photographs of excised tumors collected at endpoint. (d) Tumor growth curves during treatment. Tumor volume was calculated using the formula (length × width 2 )/2. (e) Tumor weights measured at endpoint. (f) Immunoblot analysis of VEGFR2 expression in tumor lysates from different treatment groups, β-actin was used as a reference protein. (g) IHC staining of CD31 in tumor sections from different treatment groups. Scale bar, 100 μm. (h) H&E staining of major organs (heart, liver, spleen, lung, kidney) and tumor tissues. (i) Serum ALT and AST levels measured at endpoint. Data are presented as mean ± SEM. Statistical analysis was performed by one-way ANOVA with Tukey's multiple comparisons test, ns indicates not significant, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗∗p < 0.0001.

Techniques: In Vivo, Western Blot, Expressing, Immunohistochemistry, Staining

hfNCSC-sEVs are taken up by PCs in vitro and enhance their proliferation and migration. (A) Primary cultures of hfNCSCs were established from male Sprague–Dawley rats. (B) Immunofluorescence staining of the neural crest cell marker p75 (red) and the stem cell marker nestin (green) in hfNCSCs, with 4′,6-diamidino-2-phenylindole (DAPI) staining indicating the nuclei. (C) Western blot analysis demonstrated the presence of surface markers (cluster of differentiation [CD]9, CD81, and tumor susceptibility gene 101 protein [TSG101]) and the absence of an endoplasmic reticulum marker (calnexin) in hfNCSC-sEVs. (D) Nanoparticle tracking analysis was used to quantify the concentration and size distribution of hfNCSC-sEVs. (E) Transmission electron microscopy was used to visualize the characteristic morphology of hfNCSC-sEVs. (F) Immunofluorescence staining indicated that the third-generation PCs cultured in vitro were positive for claudin-1, zonula occludens 1 (ZO1), and glucose transporter 1 (GLUT1) but negative for S100, with DAPI staining marking the nuclei. (G) The internalization of PKH26-labeled hfNCSC-sEVs (red) by ZO1-positive PCs (green) was visualized using immunofluorescence staining, with DAPI staining to mark the nuclei. (H) The Cell Counting Kit-8 assay was used to evaluate the cell viability of PCs across concentrations of 0, 2 × 10 8 , 5 × 10 8 , and 10 × 10 8 particles/mL hfNCSC-sEVs at 3, 5, and 7 days of in vitro culture ( n = 5 per group). (I) The Transwell assay was used to quantify the number of migrating PCs at 6, 12, and 18 hours post-treatment with the aforementioned concentrations of hfNCSC-sEVs, in in vitro culture ( n = 6 per group). (J) Western blot and (K) statistical analyses revealed the relative protein expression levels of proliferating cell nuclear antigen (PCNA) and vimentin in PCs from the phosphate-buffered saline (PBS) and hfNCSC-sEVs groups on day 5 of in vitro culture (normalized to β-actin, n = 3 per group). Data are expressed as the mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001 (one-way analysis of variance and Tukey’s multiple comparison test for H and I; Student’s t -test for K). The data were from at least three separate and independent studies. CCK-8: Cell counting kit-8; GLUT1: glucose transporter 1; hfNCSCs: hair follicle neural crest stem cells; ns: not significant; PCNA: proliferating cell nuclear antigen; PCs: perineurial cells; sEVs: small extracellular vesicles; ZO1: zonula occludens 1.

Journal: Neural Regeneration Research

Article Title: Small extracellular vesicles derived from hair follicle neural crest stem cells enhance perineurial cell proliferation and migration via the TGF-β/SMAD/HAS2 pathway

doi: 10.4103/NRR.NRR-D-25-00127

Figure Lengend Snippet: hfNCSC-sEVs are taken up by PCs in vitro and enhance their proliferation and migration. (A) Primary cultures of hfNCSCs were established from male Sprague–Dawley rats. (B) Immunofluorescence staining of the neural crest cell marker p75 (red) and the stem cell marker nestin (green) in hfNCSCs, with 4′,6-diamidino-2-phenylindole (DAPI) staining indicating the nuclei. (C) Western blot analysis demonstrated the presence of surface markers (cluster of differentiation [CD]9, CD81, and tumor susceptibility gene 101 protein [TSG101]) and the absence of an endoplasmic reticulum marker (calnexin) in hfNCSC-sEVs. (D) Nanoparticle tracking analysis was used to quantify the concentration and size distribution of hfNCSC-sEVs. (E) Transmission electron microscopy was used to visualize the characteristic morphology of hfNCSC-sEVs. (F) Immunofluorescence staining indicated that the third-generation PCs cultured in vitro were positive for claudin-1, zonula occludens 1 (ZO1), and glucose transporter 1 (GLUT1) but negative for S100, with DAPI staining marking the nuclei. (G) The internalization of PKH26-labeled hfNCSC-sEVs (red) by ZO1-positive PCs (green) was visualized using immunofluorescence staining, with DAPI staining to mark the nuclei. (H) The Cell Counting Kit-8 assay was used to evaluate the cell viability of PCs across concentrations of 0, 2 × 10 8 , 5 × 10 8 , and 10 × 10 8 particles/mL hfNCSC-sEVs at 3, 5, and 7 days of in vitro culture ( n = 5 per group). (I) The Transwell assay was used to quantify the number of migrating PCs at 6, 12, and 18 hours post-treatment with the aforementioned concentrations of hfNCSC-sEVs, in in vitro culture ( n = 6 per group). (J) Western blot and (K) statistical analyses revealed the relative protein expression levels of proliferating cell nuclear antigen (PCNA) and vimentin in PCs from the phosphate-buffered saline (PBS) and hfNCSC-sEVs groups on day 5 of in vitro culture (normalized to β-actin, n = 3 per group). Data are expressed as the mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001 (one-way analysis of variance and Tukey’s multiple comparison test for H and I; Student’s t -test for K). The data were from at least three separate and independent studies. CCK-8: Cell counting kit-8; GLUT1: glucose transporter 1; hfNCSCs: hair follicle neural crest stem cells; ns: not significant; PCNA: proliferating cell nuclear antigen; PCs: perineurial cells; sEVs: small extracellular vesicles; ZO1: zonula occludens 1.

Article Snippet: The following primary antibodies were used: rabbit monoclonal anti-proliferating cell nuclear antigen (PCNA) antibody (1:2000, Cat# 60097-1-Ig, Proteintech, Wuhan, China), rabbit monoclonal anti-vimentin antibody (1:1000, Cat# 5741, Cell Signaling Technology, Danvers, MA, USA), rabbit polyclonal anti-claudin-1 antibody (1:1000, Cat# 13050-1-AP, Proteintech), rabbit polyclonal anti-zonula occludens 1 (ZO1) antibody (1:10 000, Cat# 21773-1-AP, Proteintech), rabbit polyclonal anti-mothers against decapentaplegic homolog (SMAD)7 antibody (1:500, Cat# WL02975, Wanleibio, Shenyang, China), rabbit polyclonal anti-SMAD2/3 antibody (1:1000, Cat# WL01520, Wanleibio), rabbit polyclonal anti-p-SMAD2/3 antibody (1:500, Cat# WL02305, Wanleibio), rabbit recombinant anti-hyaluronan synthase 2 (HAS2) antibody (1:500, Cat# DF13702, Affinity, Cincinnati, OH, USA), rabbit monoclonal anti-β-actin antibody (1:1000, Cat# 4970, Cell Signaling Technology), and mouse monoclonal anti-β-tubulin antibody (1:5000, Cat# M20005 , Abmart, Shanghai, China).

Techniques: In Vitro, Migration, Immunofluorescence, Staining, Marker, Western Blot, Concentration Assay, Transmission Assay, Electron Microscopy, Cell Culture, Labeling, Cell Counting, Transwell Assay, Expressing, Saline, Comparison, CCK-8 Assay

hfNCSC-sEVs enhance tube formation and barrier function in PCs and promote tight junction protein expression. (A) Optical micrographs of the tube formation assay and (B) statistical analyses demonstrated the number of junctions and total length of tubes in PCs in both the phosphate-buffered saline (PBS) and hfNCSC-sEVs groups ( n = 5 per group). (C) Measurements of transmembrane resistance ( n = 3 per group) and (D) cell monolayer permeability assays ( n = 9 per group) indicated the barrier formation ability of PCs in both the PBS and hfNCSC-sEVs groups. (E) Western blot and (F) statistical analyses revealed the relative protein expression levels of the tight junction proteins zonula occludens 1 (ZO1) and claudin-1 in PCs from the PBS and hfNCSC-sEVs groups on day 7 of in vitro culture (normalized to β-actin, n = 3 per group). (G, H) Immunofluorescence staining (G) and statistical analyses (H) showed the integrated optical density (IOD) of ZO1 (green) and the expression of β-tubulin (red) in PCs from the PBS and hfNCSC-sEVs groups on day 7 of in vitro culture ( n = 3 per group). (I) Schematic illustration of the rat sciatic nerve defect model: a 5-mm defect was surgically created in the rat sciatic nerve, which was then bridged using a silicon tube, followed by an orthotopic injection procedure. (J) Immunofluorescence staining revealed the expression of claudin-1 (red) in the proximal end of regenerated tissue in both the PBS and hfNCSC-sEVs groups on day 7 post-operation, with 4′,6-diamidino-2-phenylindole (DAPI) staining indicating the nuclei. Data are expressed as the mean ± SEM. * P < 0.05, *** P < 0.001 (Student’s t -test for B, C, D, F, and H). The data were from at least three separate and independent studies. hfNCSCs: Hair follicle neural crest stem cells; IOD: integrated optical density; PCs: perineurial cells; sEVs: small extracellular vesicles; ZO1: zonula occludens 1.

Journal: Neural Regeneration Research

Article Title: Small extracellular vesicles derived from hair follicle neural crest stem cells enhance perineurial cell proliferation and migration via the TGF-β/SMAD/HAS2 pathway

doi: 10.4103/NRR.NRR-D-25-00127

Figure Lengend Snippet: hfNCSC-sEVs enhance tube formation and barrier function in PCs and promote tight junction protein expression. (A) Optical micrographs of the tube formation assay and (B) statistical analyses demonstrated the number of junctions and total length of tubes in PCs in both the phosphate-buffered saline (PBS) and hfNCSC-sEVs groups ( n = 5 per group). (C) Measurements of transmembrane resistance ( n = 3 per group) and (D) cell monolayer permeability assays ( n = 9 per group) indicated the barrier formation ability of PCs in both the PBS and hfNCSC-sEVs groups. (E) Western blot and (F) statistical analyses revealed the relative protein expression levels of the tight junction proteins zonula occludens 1 (ZO1) and claudin-1 in PCs from the PBS and hfNCSC-sEVs groups on day 7 of in vitro culture (normalized to β-actin, n = 3 per group). (G, H) Immunofluorescence staining (G) and statistical analyses (H) showed the integrated optical density (IOD) of ZO1 (green) and the expression of β-tubulin (red) in PCs from the PBS and hfNCSC-sEVs groups on day 7 of in vitro culture ( n = 3 per group). (I) Schematic illustration of the rat sciatic nerve defect model: a 5-mm defect was surgically created in the rat sciatic nerve, which was then bridged using a silicon tube, followed by an orthotopic injection procedure. (J) Immunofluorescence staining revealed the expression of claudin-1 (red) in the proximal end of regenerated tissue in both the PBS and hfNCSC-sEVs groups on day 7 post-operation, with 4′,6-diamidino-2-phenylindole (DAPI) staining indicating the nuclei. Data are expressed as the mean ± SEM. * P < 0.05, *** P < 0.001 (Student’s t -test for B, C, D, F, and H). The data were from at least three separate and independent studies. hfNCSCs: Hair follicle neural crest stem cells; IOD: integrated optical density; PCs: perineurial cells; sEVs: small extracellular vesicles; ZO1: zonula occludens 1.

Techniques: Expressing, Tube Formation Assay, Saline, Permeability, Western Blot, In Vitro, Immunofluorescence, Staining, Injection

miR-21-5p activates the TGF-β/SMAD pathway by inhibiting SMAD7, resulting in upregulated HAS2 and increased PC proliferation and migration. (A, B) The volcano plot (A) and heatmap (B) illustrated the differential gene expression in regenerated tissue between the phosphate-buffered saline (PBS) and small extracellular vesicles derived from hair follicle neural crest stem cells (hfNCSC-sEVs) groups on day 7 post-operation ( n = 3 per group). (C) The chord diagram of the Gene Ontology enrichment analysis and (D) bubble chart of the Kyoto Encyclopedia of Genes and Genomes enrichment analysis for differential genes highlighted the enriched functions and associated pathways. (E) The pie chart displayed the top 10 most abundant microRNAs (miRNAs) in hfNCSC-sEVs. (F) Schematic representation of the predicted binding sites of miR-21-5p on its target gene Smad7 , as identified by TargetScan. (G) Quantitative reverse transcription-polymerase chain reaction results indicated the relative expression levels of miR-21-5p in PCs from the PBS and hfNCSC-sEVs groups on day 3 of in vitro culture ( n = 3 per group). (H) The dual-luciferase reporter assay demonstrated the relative luciferase activity following the co-transfection of reporter constructs from the Control, wild-type (WT) Smad7 3′ untranslated region (UTR), and mutant Smad7 3′ UTR groups with either miR-21-5p overexpression plasmid or negative control vector ( n = 3 per group). (I, J) Western blot (I) and statistical analysis (J) revealed the relative protein expression levels of SMAD7 and HAS2, as well as the phosphorylation levels of SMAD2/3 in PCs from the –/–, TGF-β/–, and TGF-β/miR-21-5p groups on day 5 of in vitro culture (normalized to β-actin, n = 3 per group). Data are expressed as the mean ± SEM. ** P < 0.01, *** P < 0.001 (Student’s t -test for G; one-way analysis of variance and Tukey’s multiple comparison test for J, H). The data were from at least three separate and independent studies. GO: Gene Ontology; HAS2: hyaluronan synthase 2; hfNCSCs: hair follicle neural crest stem cells; KEGG: Kyoto Encyclopedia of Genes and Genomes; MUT: mutant; NC: negative control; ns: not significant; PCs: perineurial cells; sEVs: small extracellular vesicles; SMAD: mothers against decapentaplegic homolog; TGF-β: transforming growth factor-beta; WT: wild type.

Journal: Neural Regeneration Research

Article Title: Small extracellular vesicles derived from hair follicle neural crest stem cells enhance perineurial cell proliferation and migration via the TGF-β/SMAD/HAS2 pathway

doi: 10.4103/NRR.NRR-D-25-00127

Figure Lengend Snippet: miR-21-5p activates the TGF-β/SMAD pathway by inhibiting SMAD7, resulting in upregulated HAS2 and increased PC proliferation and migration. (A, B) The volcano plot (A) and heatmap (B) illustrated the differential gene expression in regenerated tissue between the phosphate-buffered saline (PBS) and small extracellular vesicles derived from hair follicle neural crest stem cells (hfNCSC-sEVs) groups on day 7 post-operation ( n = 3 per group). (C) The chord diagram of the Gene Ontology enrichment analysis and (D) bubble chart of the Kyoto Encyclopedia of Genes and Genomes enrichment analysis for differential genes highlighted the enriched functions and associated pathways. (E) The pie chart displayed the top 10 most abundant microRNAs (miRNAs) in hfNCSC-sEVs. (F) Schematic representation of the predicted binding sites of miR-21-5p on its target gene Smad7 , as identified by TargetScan. (G) Quantitative reverse transcription-polymerase chain reaction results indicated the relative expression levels of miR-21-5p in PCs from the PBS and hfNCSC-sEVs groups on day 3 of in vitro culture ( n = 3 per group). (H) The dual-luciferase reporter assay demonstrated the relative luciferase activity following the co-transfection of reporter constructs from the Control, wild-type (WT) Smad7 3′ untranslated region (UTR), and mutant Smad7 3′ UTR groups with either miR-21-5p overexpression plasmid or negative control vector ( n = 3 per group). (I, J) Western blot (I) and statistical analysis (J) revealed the relative protein expression levels of SMAD7 and HAS2, as well as the phosphorylation levels of SMAD2/3 in PCs from the –/–, TGF-β/–, and TGF-β/miR-21-5p groups on day 5 of in vitro culture (normalized to β-actin, n = 3 per group). Data are expressed as the mean ± SEM. ** P < 0.01, *** P < 0.001 (Student’s t -test for G; one-way analysis of variance and Tukey’s multiple comparison test for J, H). The data were from at least three separate and independent studies. GO: Gene Ontology; HAS2: hyaluronan synthase 2; hfNCSCs: hair follicle neural crest stem cells; KEGG: Kyoto Encyclopedia of Genes and Genomes; MUT: mutant; NC: negative control; ns: not significant; PCs: perineurial cells; sEVs: small extracellular vesicles; SMAD: mothers against decapentaplegic homolog; TGF-β: transforming growth factor-beta; WT: wild type.

Techniques: Migration, Gene Expression, Saline, Derivative Assay, Binding Assay, Reverse Transcription, Polymerase Chain Reaction, Expressing, In Vitro, Luciferase, Reporter Assay, Activity Assay, Cotransfection, Construct, Control, Mutagenesis, Over Expression, Plasmid Preparation, Negative Control, Western Blot, Phospho-proteomics, Comparison

miR-21-5p in hfNCSC-sEVs augments cell proliferation and migration by enhancing HAS2 expression in PCs. (A, B) Western blot (A) and statistical analyses (B) revealed the relative protein expression levels of HAS2, proliferating cell nuclear antigen (PCNA), and vimentin in PCs across the –/–, –/si- Has2 , hfNCSC-sEVs/–, and hfNCSC-sEVs/si- Has2 groups on day 5 of in vitro culture (normalized to β-actin, n = 3 per group). (C, D) The wound healing assay (C) and statistical analysis (D) demonstrated the migration rates of PCs in the aforementioned groups ( n = 3 per group). (E) The Cell Counting Kit-8 assay was used to assess cell viability in PCs across the same groups on day 5 of in vitro culture ( n = 5 per group). (F, G) Western blot (F) and statistical analyses (G) indicated the relative protein expression levels of HAS2, PCNA, and vimentin in PCs treated with phosphate-buffered saline (PBS), hfNCSC-sEVs, or hfNCSC-sEVs + miR-21-5p inhibitor on day 5 of in vitro culture (normalized to β-actin, n = 3 per group). (H–J) Immunofluorescence staining visualized the expression of HAS2 (red) and 5-ethynyl-2′-deoxyuridine (EdU; green) in PCs (H), and statistical analysis revealed the integrated optical density (IOD) of zonula occludens 1 (ZO1; I) and the cell proliferation rates (J) in the PBS, hfNCSC-sEVs, and hfNCSC-sEVs + miR-21-5p inhibitor groups on day 5 of in vitro culture ( n = 3 per group). (K, L) Western blot (K) and statistical analyses (L) showed the relative protein expression levels of HAS2, PCNA, and vimentin in regenerated tissue from the PBS, hfNCSC-sEVs, and hfNCSC-sEVs + miR-21-5p inhibitor groups on day 5 post-operation (normalized to β-tubulin, n = 3 per group). Data are expressed as the mean ± SEM. ** P < 0.01, *** P < 0.001 (one-way analysis of variance and Tukey’s multiple comparison test for B, D, E, G, I, J, and L). The data were from at least three separate and independent studies. CCK-8: Cell counting kit-8; EdU: 5-ethynyl-2′-deoxyuridine; HAS2: hyaluronan synthase 2; hfNCSCs: hair follicle neural crest stem cells; IOD: integrated optical density; PCNA: proliferating cell nuclear antigen; PCs: perineurial cells; sEVs: small extracellular vesicles; ZO1: zonula occludens 1.

Journal: Neural Regeneration Research

Article Title: Small extracellular vesicles derived from hair follicle neural crest stem cells enhance perineurial cell proliferation and migration via the TGF-β/SMAD/HAS2 pathway

doi: 10.4103/NRR.NRR-D-25-00127

Figure Lengend Snippet: miR-21-5p in hfNCSC-sEVs augments cell proliferation and migration by enhancing HAS2 expression in PCs. (A, B) Western blot (A) and statistical analyses (B) revealed the relative protein expression levels of HAS2, proliferating cell nuclear antigen (PCNA), and vimentin in PCs across the –/–, –/si- Has2 , hfNCSC-sEVs/–, and hfNCSC-sEVs/si- Has2 groups on day 5 of in vitro culture (normalized to β-actin, n = 3 per group). (C, D) The wound healing assay (C) and statistical analysis (D) demonstrated the migration rates of PCs in the aforementioned groups ( n = 3 per group). (E) The Cell Counting Kit-8 assay was used to assess cell viability in PCs across the same groups on day 5 of in vitro culture ( n = 5 per group). (F, G) Western blot (F) and statistical analyses (G) indicated the relative protein expression levels of HAS2, PCNA, and vimentin in PCs treated with phosphate-buffered saline (PBS), hfNCSC-sEVs, or hfNCSC-sEVs + miR-21-5p inhibitor on day 5 of in vitro culture (normalized to β-actin, n = 3 per group). (H–J) Immunofluorescence staining visualized the expression of HAS2 (red) and 5-ethynyl-2′-deoxyuridine (EdU; green) in PCs (H), and statistical analysis revealed the integrated optical density (IOD) of zonula occludens 1 (ZO1; I) and the cell proliferation rates (J) in the PBS, hfNCSC-sEVs, and hfNCSC-sEVs + miR-21-5p inhibitor groups on day 5 of in vitro culture ( n = 3 per group). (K, L) Western blot (K) and statistical analyses (L) showed the relative protein expression levels of HAS2, PCNA, and vimentin in regenerated tissue from the PBS, hfNCSC-sEVs, and hfNCSC-sEVs + miR-21-5p inhibitor groups on day 5 post-operation (normalized to β-tubulin, n = 3 per group). Data are expressed as the mean ± SEM. ** P < 0.01, *** P < 0.001 (one-way analysis of variance and Tukey’s multiple comparison test for B, D, E, G, I, J, and L). The data were from at least three separate and independent studies. CCK-8: Cell counting kit-8; EdU: 5-ethynyl-2′-deoxyuridine; HAS2: hyaluronan synthase 2; hfNCSCs: hair follicle neural crest stem cells; IOD: integrated optical density; PCNA: proliferating cell nuclear antigen; PCs: perineurial cells; sEVs: small extracellular vesicles; ZO1: zonula occludens 1.

Techniques: Migration, Expressing, Western Blot, In Vitro, Wound Healing Assay, Cell Counting, Saline, Immunofluorescence, Staining, Comparison, CCK-8 Assay

miR-21-5p in hfNCSC-sEVs enhances tight junction protein expression in PCs. (A, B) Immunofluorescence staining (A) and statistical analysis (B) demonstrated IOD of ZO1 (green) and the expression of β-tubulin (red) in PCs across the PBS, hfNCSC-sEVs, and hfNCSC-sEVs + miR-21-5p inhibitor groups on day 7 of in vitro culture ( n = 3 per group). (C) Western blot and (D) statistical analyses revealed the relative protein expression levels of the tight junction proteins ZO1 and claudin-1 in PCs from the PBS, hfNCSC-sEVs, and hfNCSC-sEVs + miR-21-5p inhibitor groups on day 7 of in vitro culture (normalized to β-actin, n = 3 per group). (E) Immunofluorescence staining depicted the expression of claudin-1 (red) at the proximal end of regenerated tissue in the PBS, hfNCSC-sEVs, and hfNCSC-sEVs + miR-21-5p inhibitor groups on day 7 post-operation, with DAPI staining highlighting the nuclei. (F, G) Western blot (F) and statistical analyses (G) indicated the relative protein expression levels of ZO1 and claudin-1 in regenerated tissue across the PBS, hfNCSC-sEVs, and hfNCSC-sEVs + miR-21-5p inhibitor groups on day 7 post-operation (normalized to β-actin, n = 3 per group). Data are expressed as the mean ± SEM. ** P < 0.01, *** P < 0.001 (one-way analysis of variance and Tukey’s multiple comparison test for B, D, and G). The data were from at least three separate and independent studies. DAPI: 4,6-Diamidino-2-phenylindole; hfNCSCs: hair follicle neural crest stem cells; IOD: integrated optical density; PBS: phosphate-buffered saline; PCs: perineurial cells; sEVs: small extracellular vesicles; ZO1: zonula occludens 1.

Journal: Neural Regeneration Research

Article Title: Small extracellular vesicles derived from hair follicle neural crest stem cells enhance perineurial cell proliferation and migration via the TGF-β/SMAD/HAS2 pathway

doi: 10.4103/NRR.NRR-D-25-00127

Figure Lengend Snippet: miR-21-5p in hfNCSC-sEVs enhances tight junction protein expression in PCs. (A, B) Immunofluorescence staining (A) and statistical analysis (B) demonstrated IOD of ZO1 (green) and the expression of β-tubulin (red) in PCs across the PBS, hfNCSC-sEVs, and hfNCSC-sEVs + miR-21-5p inhibitor groups on day 7 of in vitro culture ( n = 3 per group). (C) Western blot and (D) statistical analyses revealed the relative protein expression levels of the tight junction proteins ZO1 and claudin-1 in PCs from the PBS, hfNCSC-sEVs, and hfNCSC-sEVs + miR-21-5p inhibitor groups on day 7 of in vitro culture (normalized to β-actin, n = 3 per group). (E) Immunofluorescence staining depicted the expression of claudin-1 (red) at the proximal end of regenerated tissue in the PBS, hfNCSC-sEVs, and hfNCSC-sEVs + miR-21-5p inhibitor groups on day 7 post-operation, with DAPI staining highlighting the nuclei. (F, G) Western blot (F) and statistical analyses (G) indicated the relative protein expression levels of ZO1 and claudin-1 in regenerated tissue across the PBS, hfNCSC-sEVs, and hfNCSC-sEVs + miR-21-5p inhibitor groups on day 7 post-operation (normalized to β-actin, n = 3 per group). Data are expressed as the mean ± SEM. ** P < 0.01, *** P < 0.001 (one-way analysis of variance and Tukey’s multiple comparison test for B, D, and G). The data were from at least three separate and independent studies. DAPI: 4,6-Diamidino-2-phenylindole; hfNCSCs: hair follicle neural crest stem cells; IOD: integrated optical density; PBS: phosphate-buffered saline; PCs: perineurial cells; sEVs: small extracellular vesicles; ZO1: zonula occludens 1.

Techniques: Expressing, Immunofluorescence, Staining, In Vitro, Western Blot, Comparison, Saline

Signaling pathways of Garrya flavescens extract under LPS treatment in microglia. ( A ) Western blot analysis of BV-2 cells pre-treated with vehicle or GF, followed by LPS treatment at 10, 30 and 60 min. Antibodies against phosphorylated ERK, p38, AKT and β-Actin were used. ( B – D ) Quantitative analysis of the band intensities: p-ERK/β-Actin ( B ), p-p38/β-Actin ( C ), and p-AKT/β-Actin ( D ). * p < 0.05; ** p < 0.01; Student’s t -test. N = 3 independent cultures. Bars indicate mean ± SD. Abbreviations: GF, Garrya flavescens ; LPS, lipopolysaccharide; CGA, chlorogenic acid; ERK, extracellular signal-regulated kinase; p38, p38 mitogen-activated protein kinase; AKT, protein kinase B; β-actin, beta-actin.

Journal: Plants

Article Title: Anti-Inflammatory Properties of Garrya flavescens : Phytochemical Profiling and Mitigation of LPS-Induced Neuroinflammation via ERK Signaling and Mitochondrial Modulation

doi: 10.3390/plants15091319

Figure Lengend Snippet: Signaling pathways of Garrya flavescens extract under LPS treatment in microglia. ( A ) Western blot analysis of BV-2 cells pre-treated with vehicle or GF, followed by LPS treatment at 10, 30 and 60 min. Antibodies against phosphorylated ERK, p38, AKT and β-Actin were used. ( B – D ) Quantitative analysis of the band intensities: p-ERK/β-Actin ( B ), p-p38/β-Actin ( C ), and p-AKT/β-Actin ( D ). * p < 0.05; ** p < 0.01; Student’s t -test. N = 3 independent cultures. Bars indicate mean ± SD. Abbreviations: GF, Garrya flavescens ; LPS, lipopolysaccharide; CGA, chlorogenic acid; ERK, extracellular signal-regulated kinase; p38, p38 mitogen-activated protein kinase; AKT, protein kinase B; β-actin, beta-actin.

Article Snippet: Rabbit anti-β-actin antibody (bs-0061R) was obtained from Bioss (Beijing, China).

Techniques: Protein-Protein interactions, Western Blot

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