Staining:
Article Title: The Drp1-CoQ10-Coa6-ETC axis represents a therapeutic potential for working memory impairment caused by neuronal mitochondrial dysfunction
Article Snippet: After three washes with PBS, the sections were incubated for 2 h with secondary antibodies, including DyLight 647 goat anti-mouse IgG (1:5000, A23610, Abbkine, Wuhan, China) or DyLight 647 goat anti-rabbit IgG (1:5000, A23620, Abbkine). .. The sections were washed three times with PBS, and cell nuclei were stained with DAPI (1:2500, C1002, Beyotime, Shanghai, China) for 5 min. After air drying, the slides were sealed with a fluorescent mounting medium and observed using a confocal laser scanning microscope (Leica STELLARIS5). ..
Article Title: Targeted delivery of lupeol via hyaluronic acid-modified ZIF-8 for anti-hepatic fibrosis therapy
Article Snippet: Quantitative analysis was performed using flow cytometry (Beckman Coulter, USA). .. A DAPI Staining Solution (Beyotime Biotechnology, Shanghai, China) was used to perform the cell apoptosis shape assay on LX-2 cells according to the manufacturer's instructions. .. Cells were seeded into six-well plates at a density of 1.0 × 10 6 cells per well and cultured for 24 h. The next day, cells were treated with different media containing Lupeol, Lupeol@ZIF-8 and HA/Lupeol@ZIF-8 (60 μM lupeol), with blank and model groups included as controls, and incubated for an additional 24 h. For qualitative analysis, DAPI solution was added 24 h after treatment, and fluorescence signals were recorded using a fluorescence microscope (Olympus, Tokyo, Japan).
Article Title: Geometry-driven immunomodulation in 3D-printed bioceramics: Negative curvature promotes macrophage M2 polarization via Ras-MAPK/HIF-1α signaling for vascularized osteogenesis
Article Snippet: The cells types utilized in this study included: hBMSCs (No. 7500, ScienCell, USA.), Raw264.7 (CL-0190, Procell system, China), HUVECs (No. 8000, ScienCell, USA.), rBMSCs (MUBMX-01001, OriCell, USA.) and MS1(CL-0162, Procell system, China). .. The reagents used in the experiment included: H-DMEM(11965118, Gibco, USA.), α-DMEM medium(12571063, Gibco, USA.), TritonX-100(ST1723, Beyotime, China), 4 % paraformaldehyde (BL539A, Biosharp, China),FBS(A5256701, Gibco, USA.),ECM medium (Science Cell, USA.),and DAPI staining solution (C1006, Beyotime, China),BCIP/NBT(C3206, Beyotime, China), reactive oxygen species kit (S0033S, Beyotime, China), BSA (B2064, ≥98 %, Sigma-Aldrich, USA.),CD31 antibody (ab28364, Abcam, USA.), secondary anti-IGg (ab175773, Alexa Fluor® 680, Abcam, USA.), Phalloidin-iFluor 488(ab176753, Abcam, USA.), CCR7(AF5293, Bioss, China), CD206 (bsm-60761R, Bioss, China), iNOS (bs-22924R, Bioss, China), RIPA (P0013, Beyotime, China), p-ERK1/2 (AF3687, Affinity, USA.) and ERK1/2 (#AF0155, Affinity, USA), luminol detection reagent (sc-2048, Santa Cruz, USA.), GAPDH (Cat#KC-5G5, Kangchen Biotechnology, China), Trizol(15596026CN, Invitrogen, USA.), DEPC(R0601, Thermo Scientific, USA.), TBST(R017R.0000, Thermo Scientific, USA.), HIF-1a(GTX127309, GeneTex, USA.), β-Tubulin(10094-1-AP, Proteintech, UK) Adezmapimod (SB 203580, MCE, USA.) medium and Paclitaxel (99.88 %, HY-B0015R, MCE), ELISA Arg-1(E-EL-M3092, ELabSci@, China), TNF-α (E-EL-M3063, ELabSci@, China), OPN(22952-1-AP, Proteintech, UK.), F4/80 (GB11027-100, Servicebio, China) Alkaline phosphatase activity kit (P0321S, Beyotime, China), Matrigel (CLS356234, Corning, USA), Microfill MV120 (Flow tech, USA), EDTA(17892, Thermo Scientific, USA.) xylene(X112051, AR,99 %, Aladdin, China), ethanol (107-21-1, AR,99 %, Aladdin, China). .. All primers were designed and synthesized by Genema (Shanghai) Co., Ltd. 5.2.Fabrication of high-throughput array cell culture chip (HCGC) with Gaussian curvature gradient distribution. (1) Constructing models with different Gaussian curvatures: the multipipe of Grasshopper in Rhinocero software is used to regulate Gaussian curvature of the truss in the diamond framework in .
Article Title: Nucleoporin TPR integrates MAPK signaling with mitogen-induced transcriptional programs.
Article Snippet: .. Coverslips were then incubated with secondary antibodies at room temperature for 1 h, washed once with 1× PBS for 5 min, stained with DAPI (Beyotime, #C1005) for 5 min, washed twice with 1× PBS for 5 min, and finally mounted using 2 μL of antifade mounting medium (Beyotime, #P0126). .. The following primary antibodies were used: TPR total (Sigma-Aldrich, #HPA019661, rabbit, 1:500), TPR total (Abcam, #ab84516, rabbit, 1:500), p-TPR Ser2155 (developed in-house, mouse, 1:1000).
Article Title: Small extracellular vesicle-integrated by herbal hydrogels for spatiotemporal immunomodulation and neurovascular repair following traumatic brain injury
Article Snippet: .. The following were used: Nissl staining (G1036, Servicebio, China), One Step TUNEL Apoptosis Assay Kit (MA0224, Meilunbio, China), anti-Fluorescence Quencher (Contains DAPI) (P0131, Beyotime Biotechnology, China), Polymer-HRP Anti-Rabbit Secondary Antibody Kit (AFIHC003, AiFang biological, China), DAB Chromogenic Reagent Kit (ZLI-9019, ZSGB-BIO, China), RaPure Total RNA Kit (MD021, Magen, China), RNA Reversing Reagent Kit (RR047A, TaKaRa, Japan), ChamQTM Universal SYBR® qPCR Master Mix (Q711, Vazyme, China), Polymerase Chain Reaction Anti-Mouse IgG-HRP Kit (SV0001, Boster Biological Technology, China), Mouse IL-6 MPCLIA Kit (GLM0004, Boster Biological Technology, China) Isolation and purification of sEV by differential ultracentrifugation. ..
Article Title: Development of perilla essential oil-based nanoemulsions as anti-psoriatic and dermal delivery platforms for topical therapy
Article Snippet: PEG-40 stearyl ether (S40) was obtained from Croda GmbH (Nett et al., Germany). .. DAPI staining solution, RIPA buffer, hematoxylin and eosin (H&E) staining kit, and bicinchoninic acid (BCA) protein assay kits were obtained from Beyotime Biotechnology (Shanghai, China). .. Antibodies against Ki67, TNF-α, IL-6, IL-1β, IL-17 and a one-step TUNEL assay kit were from Servicebio (Wuhan, China).
ROS Assay:
Membrane:
Laser-Scanning Microscopy:Article Title: The Drp1-CoQ10-Coa6-ETC axis represents a therapeutic potential for working memory impairment caused by neuronal mitochondrial dysfunction
Article Snippet: After three washes with PBS, the sections were incubated for 2 h with secondary antibodies, including DyLight 647 goat anti-mouse IgG (1:5000, A23610, Abbkine, Wuhan, China) or DyLight 647 goat anti-rabbit IgG (1:5000, A23620, Abbkine). .. The sections were washed three times with PBS, and cell nuclei were stained with DAPI (1:2500, C1002, Beyotime, Shanghai, China) for 5 min. After air drying, the slides were sealed with a fluorescent mounting medium and observed using a confocal laser scanning microscope (Leica STELLARIS5). ..
Enzyme-linked Immunosorbent Assay:Article Title: Geometry-driven immunomodulation in 3D-printed bioceramics: Negative curvature promotes macrophage M2 polarization via Ras-MAPK/HIF-1α signaling for vascularized osteogenesis
Article Snippet: The cells types utilized in this study included: hBMSCs (No. 7500, ScienCell, USA.), Raw264.7 (CL-0190, Procell system, China), HUVECs (No. 8000, ScienCell, USA.), rBMSCs (MUBMX-01001, OriCell, USA.) and MS1(CL-0162, Procell system, China). .. The reagents used in the experiment included: H-DMEM(11965118, Gibco, USA.), α-DMEM medium(12571063, Gibco, USA.), TritonX-100(ST1723, Beyotime, China), 4 % paraformaldehyde (BL539A, Biosharp, China),FBS(A5256701, Gibco, USA.),ECM medium (Science Cell, USA.),and DAPI staining solution (C1006, Beyotime, China),BCIP/NBT(C3206, Beyotime, China), reactive oxygen species kit (S0033S, Beyotime, China), BSA (B2064, ≥98 %, Sigma-Aldrich, USA.),CD31 antibody (ab28364, Abcam, USA.), secondary anti-IGg (ab175773, Alexa Fluor® 680, Abcam, USA.), Phalloidin-iFluor 488(ab176753, Abcam, USA.), CCR7(AF5293, Bioss, China), CD206 (bsm-60761R, Bioss, China), iNOS (bs-22924R, Bioss, China), RIPA (P0013, Beyotime, China), p-ERK1/2 (AF3687, Affinity, USA.) and ERK1/2 (#AF0155, Affinity, USA), luminol detection reagent (sc-2048, Santa Cruz, USA.), GAPDH (Cat#KC-5G5, Kangchen Biotechnology, China), Trizol(15596026CN, Invitrogen, USA.), DEPC(R0601, Thermo Scientific, USA.), TBST(R017R.0000, Thermo Scientific, USA.), HIF-1a(GTX127309, GeneTex, USA.), β-Tubulin(10094-1-AP, Proteintech, UK) Adezmapimod (SB 203580, MCE, USA.) medium and Paclitaxel (99.88 %, HY-B0015R, MCE), ELISA Arg-1(E-EL-M3092, ELabSci@, China), TNF-α (E-EL-M3063, ELabSci@, China), OPN(22952-1-AP, Proteintech, UK.), F4/80 (GB11027-100, Servicebio, China) Alkaline phosphatase activity kit (P0321S, Beyotime, China), Matrigel (CLS356234, Corning, USA), Microfill MV120 (Flow tech, USA), EDTA(17892, Thermo Scientific, USA.) xylene(X112051, AR,99 %, Aladdin, China), ethanol (107-21-1, AR,99 %, Aladdin, China). .. All primers were designed and synthesized by Genema (Shanghai) Co., Ltd. 5.2.Fabrication of high-throughput array cell culture chip (HCGC) with Gaussian curvature gradient distribution. (1) Constructing models with different Gaussian curvatures: the multipipe of Grasshopper in Rhinocero software is used to regulate Gaussian curvature of the truss in the diamond framework in .
Activity Assay:Article Title: Geometry-driven immunomodulation in 3D-printed bioceramics: Negative curvature promotes macrophage M2 polarization via Ras-MAPK/HIF-1α signaling for vascularized osteogenesis
Article Snippet: The cells types utilized in this study included: hBMSCs (No. 7500, ScienCell, USA.), Raw264.7 (CL-0190, Procell system, China), HUVECs (No. 8000, ScienCell, USA.), rBMSCs (MUBMX-01001, OriCell, USA.) and MS1(CL-0162, Procell system, China). .. The reagents used in the experiment included: H-DMEM(11965118, Gibco, USA.), α-DMEM medium(12571063, Gibco, USA.), TritonX-100(ST1723, Beyotime, China), 4 % paraformaldehyde (BL539A, Biosharp, China),FBS(A5256701, Gibco, USA.),ECM medium (Science Cell, USA.),and DAPI staining solution (C1006, Beyotime, China),BCIP/NBT(C3206, Beyotime, China), reactive oxygen species kit (S0033S, Beyotime, China), BSA (B2064, ≥98 %, Sigma-Aldrich, USA.),CD31 antibody (ab28364, Abcam, USA.), secondary anti-IGg (ab175773, Alexa Fluor® 680, Abcam, USA.), Phalloidin-iFluor 488(ab176753, Abcam, USA.), CCR7(AF5293, Bioss, China), CD206 (bsm-60761R, Bioss, China), iNOS (bs-22924R, Bioss, China), RIPA (P0013, Beyotime, China), p-ERK1/2 (AF3687, Affinity, USA.) and ERK1/2 (#AF0155, Affinity, USA), luminol detection reagent (sc-2048, Santa Cruz, USA.), GAPDH (Cat#KC-5G5, Kangchen Biotechnology, China), Trizol(15596026CN, Invitrogen, USA.), DEPC(R0601, Thermo Scientific, USA.), TBST(R017R.0000, Thermo Scientific, USA.), HIF-1a(GTX127309, GeneTex, USA.), β-Tubulin(10094-1-AP, Proteintech, UK) Adezmapimod (SB 203580, MCE, USA.) medium and Paclitaxel (99.88 %, HY-B0015R, MCE), ELISA Arg-1(E-EL-M3092, ELabSci@, China), TNF-α (E-EL-M3063, ELabSci@, China), OPN(22952-1-AP, Proteintech, UK.), F4/80 (GB11027-100, Servicebio, China) Alkaline phosphatase activity kit (P0321S, Beyotime, China), Matrigel (CLS356234, Corning, USA), Microfill MV120 (Flow tech, USA), EDTA(17892, Thermo Scientific, USA.) xylene(X112051, AR,99 %, Aladdin, China), ethanol (107-21-1, AR,99 %, Aladdin, China). .. All primers were designed and synthesized by Genema (Shanghai) Co., Ltd. 5.2.Fabrication of high-throughput array cell culture chip (HCGC) with Gaussian curvature gradient distribution. (1) Constructing models with different Gaussian curvatures: the multipipe of Grasshopper in Rhinocero software is used to regulate Gaussian curvature of the truss in the diamond framework in .
Incubation:Article Title: Nucleoporin TPR integrates MAPK signaling with mitogen-induced transcriptional programs.
Article Snippet: .. Coverslips were then incubated with secondary antibodies at room temperature for 1 h, washed once with 1× PBS for 5 min, stained with DAPI (Beyotime, #C1005) for 5 min, washed twice with 1× PBS for 5 min, and finally mounted using 2 μL of antifade mounting medium (Beyotime, #P0126). .. The following primary antibodies were used: TPR total (Sigma-Aldrich, #HPA019661, rabbit, 1:500), TPR total (Abcam, #ab84516, rabbit, 1:500), p-TPR Ser2155 (developed in-house, mouse, 1:1000).
TUNEL Assay:Article Title: Small extracellular vesicle-integrated by herbal hydrogels for spatiotemporal immunomodulation and neurovascular repair following traumatic brain injury
Article Snippet: .. The following were used: Nissl staining (G1036, Servicebio, China), One Step TUNEL Apoptosis Assay Kit (MA0224, Meilunbio, China), anti-Fluorescence Quencher (Contains DAPI) (P0131, Beyotime Biotechnology, China), Polymer-HRP Anti-Rabbit Secondary Antibody Kit (AFIHC003, AiFang biological, China), DAB Chromogenic Reagent Kit (ZLI-9019, ZSGB-BIO, China), RaPure Total RNA Kit (MD021, Magen, China), RNA Reversing Reagent Kit (RR047A, TaKaRa, Japan), ChamQTM Universal SYBR® qPCR Master Mix (Q711, Vazyme, China), Polymerase Chain Reaction Anti-Mouse IgG-HRP Kit (SV0001, Boster Biological Technology, China), Mouse IL-6 MPCLIA Kit (GLM0004, Boster Biological Technology, China) Isolation and purification of sEV by differential ultracentrifugation. ..
Apoptosis Assay:Article Title: Small extracellular vesicle-integrated by herbal hydrogels for spatiotemporal immunomodulation and neurovascular repair following traumatic brain injury
Article Snippet: .. The following were used: Nissl staining (G1036, Servicebio, China), One Step TUNEL Apoptosis Assay Kit (MA0224, Meilunbio, China), anti-Fluorescence Quencher (Contains DAPI) (P0131, Beyotime Biotechnology, China), Polymer-HRP Anti-Rabbit Secondary Antibody Kit (AFIHC003, AiFang biological, China), DAB Chromogenic Reagent Kit (ZLI-9019, ZSGB-BIO, China), RaPure Total RNA Kit (MD021, Magen, China), RNA Reversing Reagent Kit (RR047A, TaKaRa, Japan), ChamQTM Universal SYBR® qPCR Master Mix (Q711, Vazyme, China), Polymerase Chain Reaction Anti-Mouse IgG-HRP Kit (SV0001, Boster Biological Technology, China), Mouse IL-6 MPCLIA Kit (GLM0004, Boster Biological Technology, China) Isolation and purification of sEV by differential ultracentrifugation. ..
Polymer:Article Title: Small extracellular vesicle-integrated by herbal hydrogels for spatiotemporal immunomodulation and neurovascular repair following traumatic brain injury
Article Snippet: .. The following were used: Nissl staining (G1036, Servicebio, China), One Step TUNEL Apoptosis Assay Kit (MA0224, Meilunbio, China), anti-Fluorescence Quencher (Contains DAPI) (P0131, Beyotime Biotechnology, China), Polymer-HRP Anti-Rabbit Secondary Antibody Kit (AFIHC003, AiFang biological, China), DAB Chromogenic Reagent Kit (ZLI-9019, ZSGB-BIO, China), RaPure Total RNA Kit (MD021, Magen, China), RNA Reversing Reagent Kit (RR047A, TaKaRa, Japan), ChamQTM Universal SYBR® qPCR Master Mix (Q711, Vazyme, China), Polymerase Chain Reaction Anti-Mouse IgG-HRP Kit (SV0001, Boster Biological Technology, China), Mouse IL-6 MPCLIA Kit (GLM0004, Boster Biological Technology, China) Isolation and purification of sEV by differential ultracentrifugation. ..
Real-time Polymerase Chain Reaction:Article Title: Small extracellular vesicle-integrated by herbal hydrogels for spatiotemporal immunomodulation and neurovascular repair following traumatic brain injury
Article Snippet: .. The following were used: Nissl staining (G1036, Servicebio, China), One Step TUNEL Apoptosis Assay Kit (MA0224, Meilunbio, China), anti-Fluorescence Quencher (Contains DAPI) (P0131, Beyotime Biotechnology, China), Polymer-HRP Anti-Rabbit Secondary Antibody Kit (AFIHC003, AiFang biological, China), DAB Chromogenic Reagent Kit (ZLI-9019, ZSGB-BIO, China), RaPure Total RNA Kit (MD021, Magen, China), RNA Reversing Reagent Kit (RR047A, TaKaRa, Japan), ChamQTM Universal SYBR® qPCR Master Mix (Q711, Vazyme, China), Polymerase Chain Reaction Anti-Mouse IgG-HRP Kit (SV0001, Boster Biological Technology, China), Mouse IL-6 MPCLIA Kit (GLM0004, Boster Biological Technology, China) Isolation and purification of sEV by differential ultracentrifugation. ..
Polymerase Chain Reaction:Article Title: Small extracellular vesicle-integrated by herbal hydrogels for spatiotemporal immunomodulation and neurovascular repair following traumatic brain injury
Article Snippet: .. The following were used: Nissl staining (G1036, Servicebio, China), One Step TUNEL Apoptosis Assay Kit (MA0224, Meilunbio, China), anti-Fluorescence Quencher (Contains DAPI) (P0131, Beyotime Biotechnology, China), Polymer-HRP Anti-Rabbit Secondary Antibody Kit (AFIHC003, AiFang biological, China), DAB Chromogenic Reagent Kit (ZLI-9019, ZSGB-BIO, China), RaPure Total RNA Kit (MD021, Magen, China), RNA Reversing Reagent Kit (RR047A, TaKaRa, Japan), ChamQTM Universal SYBR® qPCR Master Mix (Q711, Vazyme, China), Polymerase Chain Reaction Anti-Mouse IgG-HRP Kit (SV0001, Boster Biological Technology, China), Mouse IL-6 MPCLIA Kit (GLM0004, Boster Biological Technology, China) Isolation and purification of sEV by differential ultracentrifugation. ..
Isolation:Article Title: Small extracellular vesicle-integrated by herbal hydrogels for spatiotemporal immunomodulation and neurovascular repair following traumatic brain injury
Article Snippet: .. The following were used: Nissl staining (G1036, Servicebio, China), One Step TUNEL Apoptosis Assay Kit (MA0224, Meilunbio, China), anti-Fluorescence Quencher (Contains DAPI) (P0131, Beyotime Biotechnology, China), Polymer-HRP Anti-Rabbit Secondary Antibody Kit (AFIHC003, AiFang biological, China), DAB Chromogenic Reagent Kit (ZLI-9019, ZSGB-BIO, China), RaPure Total RNA Kit (MD021, Magen, China), RNA Reversing Reagent Kit (RR047A, TaKaRa, Japan), ChamQTM Universal SYBR® qPCR Master Mix (Q711, Vazyme, China), Polymerase Chain Reaction Anti-Mouse IgG-HRP Kit (SV0001, Boster Biological Technology, China), Mouse IL-6 MPCLIA Kit (GLM0004, Boster Biological Technology, China) Isolation and purification of sEV by differential ultracentrifugation. ..
Purification:Article Title: Small extracellular vesicle-integrated by herbal hydrogels for spatiotemporal immunomodulation and neurovascular repair following traumatic brain injury
Article Snippet: .. The following were used: Nissl staining (G1036, Servicebio, China), One Step TUNEL Apoptosis Assay Kit (MA0224, Meilunbio, China), anti-Fluorescence Quencher (Contains DAPI) (P0131, Beyotime Biotechnology, China), Polymer-HRP Anti-Rabbit Secondary Antibody Kit (AFIHC003, AiFang biological, China), DAB Chromogenic Reagent Kit (ZLI-9019, ZSGB-BIO, China), RaPure Total RNA Kit (MD021, Magen, China), RNA Reversing Reagent Kit (RR047A, TaKaRa, Japan), ChamQTM Universal SYBR® qPCR Master Mix (Q711, Vazyme, China), Polymerase Chain Reaction Anti-Mouse IgG-HRP Kit (SV0001, Boster Biological Technology, China), Mouse IL-6 MPCLIA Kit (GLM0004, Boster Biological Technology, China) Isolation and purification of sEV by differential ultracentrifugation. ..
Bicinchoninic Acid Protein Assay:Article Title: Development of perilla essential oil-based nanoemulsions as anti-psoriatic and dermal delivery platforms for topical therapy
Article Snippet: PEG-40 stearyl ether (S40) was obtained from Croda GmbH (Nett et al., Germany). .. DAPI staining solution, RIPA buffer, hematoxylin and eosin (H&E) staining kit, and bicinchoninic acid (BCA) protein assay kits were obtained from Beyotime Biotechnology (Shanghai, China). .. Antibodies against Ki67, TNF-α, IL-6, IL-1β, IL-17 and a one-step TUNEL assay kit were from Servicebio (Wuhan, China).
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