Journal: International Journal of Nanomedicine

Article Title: Bone Regeneration Revolution: Pulsed Electromagnetic Field Modulates Macrophage-Derived Exosomes to Attenuate Osteoclastogenesis

doi: 10.2147/IJN.S470901

Figure Lengend Snippet: SEM analyses of THP-1: ( A and B ) in normal conditions, cells acquire a round morphology typical of M1 phenotype; meanwhile, ( C and D ) in presence of PEMF, they acquire a fusiform morphology typical of M2 phenotype. ( A and C ) Scale Bar: 200 μm, ( B and D ) Scale Bar: 30 μm.

Article Snippet: The THP-1 monocyte cell line, acquired from Resnova (Rome, Italy), underwent culture at Roswell Park Memorial Institute (RPMI) 1640 complete medium (Euroclone S.P.A, Pero, Italy) and 10% Fetal bovine serum (FBS, Euroclone S.P.A., Pero.

Techniques:

Journal: International Journal of Nanomedicine

Article Title: Bone Regeneration Revolution: Pulsed Electromagnetic Field Modulates Macrophage-Derived Exosomes to Attenuate Osteoclastogenesis

doi: 10.2147/IJN.S470901

Figure Lengend Snippet: Heat map and histogram related to gene expression of principal genes expressed by THP-1 cells in presence of PEMF.

Article Snippet: The THP-1 monocyte cell line, acquired from Resnova (Rome, Italy), underwent culture at Roswell Park Memorial Institute (RPMI) 1640 complete medium (Euroclone S.P.A, Pero, Italy) and 10% Fetal bovine serum (FBS, Euroclone S.P.A., Pero.

Techniques: Expressing

Journal: Bioscience Reports

Article Title: LncRNA GNAS-AS1 facilitates ER + breast cancer cells progression by promoting M2 macrophage polarization via regulating miR-433-3p/GATA3 axis

doi: 10.1042/BSR20200626

Figure Lengend Snippet: ( A ) The relative quantity of GNAS-AS1 in ER + breast cancer tissues and adjacent normal tissues was examined by qRT-PCR. ( B ) Human monocytes were isolated from PBMCs with antibody against CD14 and CD11b and analyzed using flow cytometry. ( C ) Flow cytometry was used to quantify the proportion of M1 or M2 macrophages. ( D ) M1 macrophage markers (TNF-α, IL-6) were measured by qRT-PCR. ( E ) M2 macrophage markers (IL-10, Arginase-1) were detected by qRT-PCR. ( F ) qRT-PCR was performed to determine GNAS-AS1 expression in TAMs (M0, M1, M2). ( G ) The expression level of GNAS-AS1 in breast cancer cells (T47D and MCF-7) and normal mammary epithelial cells (MCF10A) were determined by qRT-PCR. Data with error bars are presented as the mean ± SD; * P <0.05, ** P <0.01, *** P <0.001 as determined by the Student’s t test or one-way ANOVA test.

Article Snippet: The cells were maintained in RPMI 1640 medium (THP-1) (Hyclone, America), MEGM medium (MCF10A) (Hyclone, America) and DMEM/F-12 (T47D, MCF-7) (Gibco, America), supplemented with 10% (v/v) fetal bovine serum (FBS, Gibco, America) and 1% penicillin/streptomycin (Beyotime Biotechnology, China).

Techniques: Quantitative RT-PCR, Isolation, Flow Cytometry, Expressing

Journal: Bioscience Reports

Article Title: LncRNA GNAS-AS1 facilitates ER + breast cancer cells progression by promoting M2 macrophage polarization via regulating miR-433-3p/GATA3 axis

doi: 10.1042/BSR20200626

Figure Lengend Snippet: In IL-4 treated THP-1-differentiated macrophages, pSin-GNAS-AS1 plasmids or negative plasmids were transfected into cells, then next experiments were conducted. ( A ) The level of GNAS-AS1 was evaluated using qRT-PCR. ( B ) Flow cytometry was used to quantify the proportion of M1 or M2 macrophages. ( C ) qRT-PCR was performed to determine the expression levels of M2 macrophage markers (IL-10, Arginase-1). ( D ) CCK-8 assay was conducted to assess the cell viability of T47D and MCF-7 cells co-cultured with above treated THP-1-differentiated macrophages. ( E ) Wound healing assay was used to examine the migration of T47D and MCF-7 cells co-cultured with above treated THP-1-differentiated macrophages. ( F ) Transwell was applied to detect the invasion of T47D and MCF-7 cells co-cultured with above treated THP-1-differentiated macrophages. Data with error bars are presented as the mean ± SD. The Student’s t test and one-way ANOVA test were used to determine significance; * P <0.05, ** P <0.01, *** P <0.001.

Article Snippet: The cells were maintained in RPMI 1640 medium (THP-1) (Hyclone, America), MEGM medium (MCF10A) (Hyclone, America) and DMEM/F-12 (T47D, MCF-7) (Gibco, America), supplemented with 10% (v/v) fetal bovine serum (FBS, Gibco, America) and 1% penicillin/streptomycin (Beyotime Biotechnology, China).

Techniques: Transfection, Quantitative RT-PCR, Flow Cytometry, Expressing, CCK-8 Assay, Cell Culture, Wound Healing Assay, Migration

Journal: Bioscience Reports

Article Title: LncRNA GNAS-AS1 facilitates ER + breast cancer cells progression by promoting M2 macrophage polarization via regulating miR-433-3p/GATA3 axis

doi: 10.1042/BSR20200626

Figure Lengend Snippet: ( A ) The binding site of miR-433-3p on GNAS-AS1 3′-UTR region. ( B ) Dual luciferase reporter assay was used to validate the molecular relationship between miR-433-3p and GNAS-AS1 in THP-1-differentiated macrophages. ( C ) qRT-PCR was performed to assess the expression of GNAS-AS1 in THP-1-differentiated macrophages transfected with pSin-GNAS-AS1 plasmids or si-GNAS-AS1. ( D ) qRT-PCR was performed to assess the expression of miR-433-3p in THP-1-differentiated macrophages transfected with pSin-GNAS-AS1 plasmids or si-GNAS-AS1. Data with error bars are presented as the mean ± SD. The Student’s t test and one-way ANOVA test were used to determine significance; * P <0.05, ** P <0.01, *** P <0.001.

Article Snippet: The cells were maintained in RPMI 1640 medium (THP-1) (Hyclone, America), MEGM medium (MCF10A) (Hyclone, America) and DMEM/F-12 (T47D, MCF-7) (Gibco, America), supplemented with 10% (v/v) fetal bovine serum (FBS, Gibco, America) and 1% penicillin/streptomycin (Beyotime Biotechnology, China).

Techniques: Binding Assay, Luciferase, Reporter Assay, Quantitative RT-PCR, Expressing, Transfection

Journal: Bioscience Reports

Article Title: LncRNA GNAS-AS1 facilitates ER + breast cancer cells progression by promoting M2 macrophage polarization via regulating miR-433-3p/GATA3 axis

doi: 10.1042/BSR20200626

Figure Lengend Snippet: In IL-4 treated THP-1-differentiated macrophages, pSin-GNAS-AS1 plasmids were co-transfected with/without miR-433-3p mimics, then next experiments were conducted. ( A ) qRT-PCR was performed to analyze the expression of miR-433-3p. ( B ) Flow cytometry was used to quantify the proportion of M1 or M2 macrophages. ( C ) qRT-PCR was applied to determine the expressions of M2 macrophage markers (IL-10, Arginase-1). ( D ) CCK-8 assay was conducted to assess the cell viability of T47D and MCF-7 cells co-cultured with above treated THP-1-differentiated macrophages. ( E ) Wound healing assay was used to detect the migration of T47D and MCF-7 cells co-cultured with above treated THP-1-differentiated macrophages. ( F ) Transwell assay was performed to elevate the invasion of T47D and MCF-7 cells co-cultured with above treated THP-1-differentiated macrophages. Data with error bars are presented as the mean ± SD. The Student’s t test and one-way ANOVA test were used to determine significance; * P <0.05, ** P <0.01, *** P <0.001.

Article Snippet: The cells were maintained in RPMI 1640 medium (THP-1) (Hyclone, America), MEGM medium (MCF10A) (Hyclone, America) and DMEM/F-12 (T47D, MCF-7) (Gibco, America), supplemented with 10% (v/v) fetal bovine serum (FBS, Gibco, America) and 1% penicillin/streptomycin (Beyotime Biotechnology, China).

Techniques: Transfection, Quantitative RT-PCR, Expressing, Flow Cytometry, CCK-8 Assay, Cell Culture, Wound Healing Assay, Migration, Transwell Assay

Journal: Bioscience Reports

Article Title: LncRNA GNAS-AS1 facilitates ER + breast cancer cells progression by promoting M2 macrophage polarization via regulating miR-433-3p/GATA3 axis

doi: 10.1042/BSR20200626

Figure Lengend Snippet: ( A ) The binding site of miR-433-3p on GATA3 3′-UTR region. ( B ) Dual luciferase reporter assay was used to validate the binding relationship between miR-433-3p and GATA3 in THP-1-differentiated macrophages. ( C ) qRT-PCR was performed to assess the expression of miR-433-3p in THP-1-differentiated macrophages transfected with miR-433-3p mimics or miR-433-3p inhibitor. ( D ) qRT-PCR was performed to assess the expression of GATA3 in THP-1-differentiated macrophages transfected with miR-433-3p mimics or miR-433-3p inhibitor. Data with error bars are presented as the mean ± SD; The Student’s t test and one-way ANOVA test were used to determine significance; * P <0.05, ** P <0.01, *** P <0.001.

Article Snippet: The cells were maintained in RPMI 1640 medium (THP-1) (Hyclone, America), MEGM medium (MCF10A) (Hyclone, America) and DMEM/F-12 (T47D, MCF-7) (Gibco, America), supplemented with 10% (v/v) fetal bovine serum (FBS, Gibco, America) and 1% penicillin/streptomycin (Beyotime Biotechnology, China).

Techniques: Binding Assay, Luciferase, Reporter Assay, Quantitative RT-PCR, Expressing, Transfection

Journal: Bioscience Reports

Article Title: LncRNA GNAS-AS1 facilitates ER + breast cancer cells progression by promoting M2 macrophage polarization via regulating miR-433-3p/GATA3 axis

doi: 10.1042/BSR20200626

Figure Lengend Snippet: ( A ) The protein level of GATA3 was examined by Western blot analysis in THP-1 cells transfected with pSin-GNAS-AS1 plasmids or si-GNAS-AS1. ( B–G ) In IL-4 treated THP-1-differentiated macrophages, pSin-GNAS-AS1 plasmids were co-transfected with/without siGATA3, then, next experiments were performed. (B) The mRNA level of GATA3 was analyzed by qRT-PCR. (C) Flow cytometry was used to quantify the proportion of M2 macrophages polarization. (D) qRT-PCR was performed to determine the expressions of M2 macrophage markers (IL-10, Arginase-1). (E) CCK-8 was used to detect the migration of T47D and MCF-7 cells co-cultured with above treated THP-1-differentiated macrophages. (F) Wound healing assay was used to detect the migration of T47D and MCF-7 cells co-cultured with above treated THP-1-differentiated macrophages. (G) Transwell was used to detect the invasion of T47D and MCF-7 cells co-cultured with above treated THP-1-differentiated macrophages. Data with error bars are presented as the mean ± SD. The Student’s t test and one-way ANOVA test were used to determine significance; * P <0.05, ** P <0.01, *** P <0.001.

Article Snippet: The cells were maintained in RPMI 1640 medium (THP-1) (Hyclone, America), MEGM medium (MCF10A) (Hyclone, America) and DMEM/F-12 (T47D, MCF-7) (Gibco, America), supplemented with 10% (v/v) fetal bovine serum (FBS, Gibco, America) and 1% penicillin/streptomycin (Beyotime Biotechnology, China).

Techniques: Western Blot, Transfection, Quantitative RT-PCR, Flow Cytometry, CCK-8 Assay, Migration, Cell Culture, Wound Healing Assay

Journal: International Journal of Molecular Sciences

Article Title: Specific Targeting of STAT3 in B Cells Suppresses Progression of B Cell Lymphoma

doi: 10.3390/ijms241713666

Figure Lengend Snippet: Properties of the CSP-specific siRNA carrier system: ( A ) The graphical depiction of anti-CD19 mAb coupling to protamine by bivalent cross-linker sulfo-SMCC siRNA conjugate via electrostatic interactions. ( B ) Flow cytometric analysis showed the affinity of the CSP-FITC-labeled siRNA conjugate to B cells. Representative dot plots and summarized MFI data are shown. MFI: mean fluorescent intensity. Pseudocolor plots are a type of bivariate density plot. It displays the relative population density of cell populations within the Graph Window along two parameters. Data are shown as the mean ± SEM ( n = 3). *** p < 0.001, Student’s t -test. Red dots are from protamine:CD19 mAb at 40:1 ratio while blue dots are at 25:1 ratio. ( C ) Confocal laser scanning microscopy images of the FITC-siRNA (green), protamine FITC-siRNA, and CSP FITC-siRNA conjugate staining in B cells (B220, red). DAPI staining (blue) was used to reveal nuclei. Scale bars: 20 μm.

Article Snippet: Conjugates were then mixed with anti-mouse CD19 mAb (32 umol/L, BioXcell, Lebanon, NH, USA) in a 25:1 molar ratio at 4 °C overnight.

Techniques: Labeling, Confocal Laser Scanning Microscopy, Staining