Journal: Molecular Medicine Reports

Article Title: Characterization of bone marrow-derived mesenchymal stem cells from dimethyloxallyl glycine-preconditioned mice: Evaluation of the feasibility of dimethyloxallyl glycine as a mobilization agent

doi: 10.3892/mmr.2016.4945

Figure Lengend Snippet: Paracrine capacity of NBM-MSCs and DBM-MSCs. (A) Quantification of TGF concentration in the culture supernatant of NBM-MSCs and DBM-MSCs. There was no significant difference between the expression levels (P>0.05). (B) Quantification of PDGF concentration in the culture supernatant of NBM-MSCs and DBM-MSCs. No significant difference was identified between them (P>0.05). TGF and PDGF concentrations were detected using enzyme-linked immunosorbent assays. Data are presented as the mean ± standard deviation. BM-MSCs, bone marrow-derived mesenchymal stem cells from normal saline-treated mice; DBM-MSCs, bone marrow-derived mesenchymal stem cells from dimethyloxallyl glycine-preconditioned mice; TGF, transforming growth factor; PDGF, platelet-derived growth factor.

Article Snippet: Mouse TGF (β IG-H3) and PDGF ELISA kits (Wuhan Boster Biological Co., Ltd., Wuhan, China) were used, according to the manufacturer's protocol.

Techniques: Concentration Assay, Expressing, Standard Deviation, Derivative Assay, Saline

Journal: bioRxiv

Article Title: Hematopoietic Stem and Progenitor Cell Aging is Initiated at Middle Age Through Decline in Local Insulin-Like Growth Factor 1 (IGF1)

doi: 10.1101/2020.07.11.198846

Figure Lengend Snippet: a ) Expression pattern of Igf2 , Nrg1 , Tgfb1 , and Egf in BM subsets assessed by scRNA-seq . For detailed cell type annotation refer to: https://nicheview.shiny.embl.de . ( b ) IGF2 concentration in BM fluid of a single mouse femur from 2-4mo ( n = 7), 6mo (n = 6), 12-14mo ( n = 5) and 26-28mo ( n = 7) mice. ( c ) Expression of Igf1r in BM cells by scRNA-seq . ( d ) Left: Experimental design. Right: Frequency of donor cells (CD45.1+) and donor-derived T cells (CD45.1+ CD3+) in the blood at 24wks post-transplant of WT BM into lethally irradiated WT or Igf1 fl/fl ; CreER T2 recipient mice ( n = 5, 7, from left to right). ( e ) Left: Experimental design. Right: Frequency of donor cells (CD45.2+), and donor-derived myeloid cells (CD11b+), B cells (B220+), and T cells (CD3+) in the blood at 24wks post-transplant of WT or Igf1r fl/fl Mx1 -Cre BM into lethally irradiated WT recipient mice ( n = 7, 4, from left to right). ( f ) Left: Experimental design. Right: Frequency of donor cells (CD45.1+) and donor-derived T cells (CD45.1+ CD3+) in the blood at 20wks post-transplant of WT BM into lethally irradiated WT or Igf1 fl/fl ; Nestin-Cre ER recipient mice ( n = 6, 5, from left to right). ( g ) Top panel: Igf1 recombination PCR, middle panel: Igf1 genotyping PCR and bottom panel: Cre genotyping PCR. Input for all reactions was gDNA isolated from livers of tamoxifen-treated Igf1 +/+ or Igf1 fl/fl ; Nestin-Cre ER mice or recombined control (CNT; Igf1 fl/fl ; Cre-ER T2 ) ( n = 3, 3, from left to right). ( b, d-f ) Dots represent individual mice and bars are mean ± SEM. P -values were generated for ( b ) by one-way ANOVA with Holm-Sidak’s multiple comparisons test, ( d - f ) by unpaired, two-tailed t tests.

Article Snippet: A mouse-specific IGF1 Immunoassay kit (ELISA, R&D Systems) and a mouse-specific IGF2 PicoKine kit (ELISA, Boster biological technology) was used to determine the concentration of IGF1 and IGF2 proteins in the BM.

Techniques: Expressing, Concentration Assay, Derivative Assay, Irradiation, Isolation, Control, Generated, Two Tailed Test

Journal: Molecular Therapy. Nucleic Acids

Article Title: Novel gene therapy for drug-resistant melanoma: Synergistic combination of PTEN plasmid and BRD4 PROTAC-loaded lipid nanocarriers

doi: 10.1016/j.omtn.2024.102292

Figure Lengend Snippet: Physicochemical characteristics and in vitro tumor growth inhibitory activity of PL-NANO in VEM-resistant melanoma cell lines DLS graphs illustrating (A) unimodal particle size distribution and (B) neutral zeta potential of PL-NANO. (C) Image of fabricated PL-NANO depicting a blue tinge in the developed nanoformulation ( n = 3d Data are presented as mean ± SD). (D) Cryo-TEM image of PL-NANO reveals spherical unilamellar liposomal vesicles in the size range of 80–100 nm. (E) Representative agarose gel electrophoresis image illustrating binding efficiency and complete entrapment of PTEN plasmid to PL-NANO. (F) Cell counting assay results depicting inhibition in growth of A375V cells by PL-NANO at different time points as compared with controls. (G) Quantitative analysis of PTEN expression following transfection with PL-NANO at different doses and Transit LT1 after 48 h of treatment in A375V cells. Significant difference depicted in the graph is in comparison with controls. Control: non-treated cells ( n = 6, data are presented as mean ± SD). ∗ p < 0.05, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: Target protein expression mediated by PL-NANO was analyzed using PTEN Colorimetric Cell-Based ELISA Kit (Boster Bio, Pleasanton, CA, USA).

Techniques: In Vitro, Activity Assay, Zeta Potential Analyzer, Agarose Gel Electrophoresis, Binding Assay, Plasmid Preparation, Cell Counting, Inhibition, Expressing, Transfection, Comparison, Control

Journal: Molecular Therapy. Nucleic Acids

Article Title: Novel gene therapy for drug-resistant melanoma: Synergistic combination of PTEN plasmid and BRD4 PROTAC-loaded lipid nanocarriers

doi: 10.1016/j.omtn.2024.102292

Figure Lengend Snippet: IC 50 values of ARV in AL-NANO alone and in combination with PL-NANO in BRAFi-resistant melanoma cell lines ( n = 6, data are presented as mean ± SD)

Article Snippet: Target protein expression mediated by PL-NANO was analyzed using PTEN Colorimetric Cell-Based ELISA Kit (Boster Bio, Pleasanton, CA, USA).

Techniques: Plasmid Preparation

Journal: Molecular Therapy. Nucleic Acids

Article Title: Novel gene therapy for drug-resistant melanoma: Synergistic combination of PTEN plasmid and BRD4 PROTAC-loaded lipid nanocarriers

doi: 10.1016/j.omtn.2024.102292

Figure Lengend Snippet: Results for target protein expression, biocompatibility and in vitro hemolysis analysis for PL-NANO and AL-NANO (A) Representative western blots for expression of BRD-4, c-Myc, PTEN, MAPK, and pMAPK proteins in A375V cells are shown. β-Actin was used as an internal control. (B) Quantification of the western blot results for expression of each target protein in A375V cells. (C) The cytotoxicity curve represents negligible toxicity exhibited by PL-NANO and AL-NANO both alone and in combination in MDCK cells. PL-NANO alone resulted in more than 95% cell viability when tested in MDCK cells at a dose of 500 ng per well. Control: non-treated cells ( n = 6, data are presented as mean ± SD). (D) Negligible hemolysis demonstrated by PL-NANO at 200 μM concentration of ionizable lipid SM-102 as well as AL-NANO at concentration of 10 μg/mL ARV as tested in mice RBCs. ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Target protein expression mediated by PL-NANO was analyzed using PTEN Colorimetric Cell-Based ELISA Kit (Boster Bio, Pleasanton, CA, USA).

Techniques: Expressing, In Vitro, Western Blot, Control, Concentration Assay