Journal: Molecular Medicine Reports

Article Title: A combined antitumor strategy of separately transduced mesenchymal stem cells with soluble TRAIL and IFNβ produces a synergistic activity in the reduction of lymphoma and mice survival enlargement

doi: 10.3892/mmr.2022.12722

Figure Lengend Snippet: BM-MSCs characterization. (A) BM-MSCs were adherent to the culture flask and presented elongated morphology (scale bar, 100 µm). They also expressed high mesenchymal markers, such as (B) CD90 and (C) CD105 (scale bar, 100 µm). In addition, they could differentiate to chondrogenic by (D) Alcian blue staining and calcified areas generated by osteogenic cells with (E) Von Kossa stain. BM-MSCs, bone marrow mesenchymal stem cells.

Article Snippet: MSC multipotency was examined by inducing the cells to differentiate to osteoblasts or chondroblasts, using the appropriate reagents in the Mouse Mesenchymal Stem Cell Functional Identification kit (cat. no. SC010; R&D Systems, Inc.) and following the manufacturer's instructions.

Techniques: Staining, Generated

Journal: Molecular Medicine Reports

Article Title: A combined antitumor strategy of separately transduced mesenchymal stem cells with soluble TRAIL and IFNβ produces a synergistic activity in the reduction of lymphoma and mice survival enlargement

doi: 10.3892/mmr.2022.12722

Figure Lengend Snippet: Lentiviral transduction of BM-MSC. (A-C) Lentiviral vectors were designed to contain (A) sTRAIL, (B) flTRAIL and (C) IFNβ. (D) Representative fluorescent images of BM-MSCs transduced with lentiviral vectors; nucleus stained with DAPI (blue) and transduced BM-MSCs expressing GFP protein (green signal). (E and F) Western blot analysis confirming protein expression of (E) flTRAIL and sTRAIL (F) and IFNβ in untransfected MSCs and MSCs transduced with a lentivirus that does not contain any of the genes of interest (empty lentiviral vector pLV[Exp]-EGFP:T2A:Puro-EF1A> mCherry, Vector Builder). MSCs express each of the genes flTRAIL, sTRAIL and IFN and as positive control the recombinant protein. Actin was used as a loading control. BM-MSCs, bone marrow mesenchymal stem cells; sTRAIL, TRAIL soluble; flTRAIL, TRAIL full length; IFNβ, interferon β.

Article Snippet: MSC multipotency was examined by inducing the cells to differentiate to osteoblasts or chondroblasts, using the appropriate reagents in the Mouse Mesenchymal Stem Cell Functional Identification kit (cat. no. SC010; R&D Systems, Inc.) and following the manufacturer's instructions.

Techniques: Transduction, Staining, Expressing, Western Blot, Plasmid Preparation, Positive Control, Recombinant, Control

Journal: Molecular Medicine Reports

Article Title: A combined antitumor strategy of separately transduced mesenchymal stem cells with soluble TRAIL and IFNβ produces a synergistic activity in the reduction of lymphoma and mice survival enlargement

doi: 10.3892/mmr.2022.12722

Figure Lengend Snippet: Kaplan-Meier curves. (A) First model with all treatments. (B) Second model with sTRAIL and IFNβ treatment and MSC naïve. *P<0.05 and **P<0.001 compared with the untreated group. sTRAIL, TRAIL soluble; IFNβ, interferon β; MSC, mesenchymal stem cell.

Article Snippet: MSC multipotency was examined by inducing the cells to differentiate to osteoblasts or chondroblasts, using the appropriate reagents in the Mouse Mesenchymal Stem Cell Functional Identification kit (cat. no. SC010; R&D Systems, Inc.) and following the manufacturer's instructions.

Techniques:

Journal: Molecular cell

Article Title: Not All H3K4 methylations are Created Equal: Mll2/COMPASS Dependency in Primordial Germ Cell Specification

doi: 10.1016/j.molcel.2017.01.013

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: V6.5 mouse embryonic stem cells , Novus Biologicals , Cat#NBP1–41162.

Techniques: Recombinant, Clinical Proteomics, Purification, Knock-Out, Reporter Assay, SYBR Green Assay, Sequencing, Software, Quantitative Proteomics, Next-Generation Sequencing

Journal: PLoS Genetics

Article Title: RNAi-Dependent and Independent Control of LINE1 Accumulation and Mobility in Mouse Embryonic Stem Cells

doi: 10.1371/journal.pgen.1003791

Figure Lengend Snippet: A. Western analysis of XRN2 and L1_ORF1 accumulation in WT and Xrn2 _KD mESCs; CM: Coomassie staining of total protein. B. qRT-PCR analysis of L1_ORF2 mRNA levels in WT and Xrn2 _KD mESCs. C. qPCR analysis of L1_Tf copy-number in WT and Xrn2 _KD mESCs. D–E. qRT-PCR analysis of miR-295 (D) and L1_ORF2 mRNA (E) levels in WT and Dgcr8 _KO mESCs. F. qPCR analysis of L1_Tf copy-number in WT and Dgcr8 _KO mESCs. G–H. qRT-PCR analysis of miR-295 (G) and L1_ORF2 mRNA (H) levels upon hAgo2 deletion in Tamoxifen-treated Ago1,2,3,4_KO mESCs. I. qPCR analysis of L1_Tf copy-number in Ago1,2,3,4_KO_hAgo2 mESCs before and after hAgo2 deletion. *: p-value<0.1.

Article Snippet: TKO mESCs were described in ref. . Dgcr8 _KO mESCs were purchased from Novus Biologicals (NBA1-19349).

Techniques: Western Blot, Staining, Quantitative RT-PCR

Journal: PLoS Genetics

Article Title: RNAi-Dependent and Independent Control of LINE1 Accumulation and Mobility in Mouse Embryonic Stem Cells

doi: 10.1371/journal.pgen.1003791

Figure Lengend Snippet: A. Western analysis of endogenous AGO1, AGO2 and L1_ORF1 protein levels in Dcr Flx/Flx , Dcr −/− and Dgcr8 _KO ESCs; CM: Coomassie staining of total protein. B. Western analysis of endogenous AGO2 and L1_ORF1 protein levels in Dcr Flx/Flx , Dc r −/− mESCs and one representative stable line of h Dcr -complemented Dcr −/− mESC; CM: Coomassie staining of total protein. C–E. qRT-PCR analysis of L1_ORF2 mRNA levels (C), miR-295 and miR-16 levels (D), and Hmga2 and Btg2 mRNA levels (established targets of mmu-miR-196a and mmu-let-7a and mmu-miR-132, respectively) in the various cell lines depicted in (E).

Article Snippet: TKO mESCs were described in ref. . Dgcr8 _KO mESCs were purchased from Novus Biologicals (NBA1-19349).

Techniques: Western Blot, Staining, Quantitative RT-PCR

Journal: PLoS Genetics

Article Title: RNAi-Dependent and Independent Control of LINE1 Accumulation and Mobility in Mouse Embryonic Stem Cells

doi: 10.1371/journal.pgen.1003791

Figure Lengend Snippet: A. Visualization of Embryoid bodies from Dcr Flx/Flx , Dcr −/− and h Dcr -complemented Dcr −/− mESCs after 1, 6 and 10 days of differentiation. B. Western analysis of OCT4 and endogenous AGO2 protein levels in the cells depicted in (A) before (d0) and after 10 days of differentiation (d10); CM: Coomassie staining of total protein. C. Same as (B) but in WT, Xrn2 _KD and Dgcr8 _KO mESCs.

Article Snippet: TKO mESCs were described in ref. . Dgcr8 _KO mESCs were purchased from Novus Biologicals (NBA1-19349).

Techniques: Western Blot, Staining

Journal: Molecular cell

Article Title: Intragenic enhancers attenuate host gene expression

doi: 10.1016/j.molcel.2017.09.010

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Mouse ESCs, Dicer knock-out (KO) , Novus Biologicals , NBP1-96751.

Techniques: Recombinant, Negative Control, Lysis, Luciferase, Reporter Gene Assay, cDNA Synthesis, Gel Extraction, TA Cloning, Knock-Out, Control, Amplification, Plasmid Preparation, CRISPR, ChIP-sequencing, RNA Sequencing, Hi-C, ChIA Pet Assay

Journal: Frontiers in Immunology

Article Title: GM-CSF Inhibits c-Kit and SCF Expression by Bone Marrow-Derived Dendritic Cells

doi: 10.3389/fimmu.2017.00147

Figure Lengend Snippet: Granulocyte-macrophage colony-stimulating factor (GM-CSF) modulates c-kit and stem cell factor (SCF) expression by BM-derived DCs (BMdDCs) . (A–C) Effect of GM-CSF and cell density on c-kit expression by BMdDCs. BMdDCs were plated in 24-well plates and cultured for 2 days in complete Opti-MEM medium in four different conditions, that is at either 2 × 10 5 or 1.2 × 10 5 cell/well, and either with or without (w/o) GM-CSF at 20 ng/ml, as indicated. Cells were stained with fluorochrome-conjugated monoclonal antibodies (mAbs) and analyzed by flow cytometry. (A) Typical flow cytometric profiles, showing CD40 and MHCII expression by BMdDCs. Numbers represent percentages of cells in the indicated regions. (B) Typical histograms showing c-kit expression by MHCII int CD40 int and MHCII hi CD40 hi BMdDCs, gated as in (A) . Solid lines represent c-kit staining profiles, dashed lines represent isotype control mAb. Numbers indicate c-kit median fluorescence intensity (MFI) values. (C) Summary of c-kit expression results obtained from MHCII hi CD40 hi BMdDCs, gated as in (A) . c-kit MFI from individual samples and average values (bar). (D,E) Effect of GM-CSF on SCF expression by BMdDCs. (D) Cell lysates were prepared from day 0 BMdDCs and BMdDCs cultured for 2 days in 24-well plates at 1.2 × 10 5 cell/well in complete Opti-MEM medium with or w/o GM-CSF at 20 ng/ml, as indicated. SCF protein expression was analyzed by ELISA, testing 25 µg of cell lysate in 100 µl/well. Data are expressed as picograms per milliliter. Individual results from three experiments and average values (bar) are shown. (E) Day 0 and day 2 BMdDCs cultured in 24-well plates at 1.2 × 10 5 cell/well with or w/o GM-CSF at 20 ng/ml were analyzed by Real-Time PCR in triplicates. SCF mRNA expression was calculated relative to hprt1 in arbitrary units. For each experiment, day 2 c-kit/hprt1 levels were normalized with day 0. In (A,B) representative data of N = 4 experiments, in (C) N = 4 experiments, in (D) N = 3 experiments, in (E) mean ± SD of four experiments (* P ≤ 0.05; ** P ≤ 0.01).

Article Snippet: BMdDC culture supernatants (100 μl/well) and BMdDC lysates (25 μg of cell lysate/well) were tested by mouse SCF ELISA kit (Boster Immunoleader, Pleasanton, CA, USA).

Techniques: Expressing, Derivative Assay, Cell Culture, Staining, Bioprocessing, Flow Cytometry, Control, Fluorescence, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction