Journal: Autophagy

Article Title: Kit-mediated autophagy suppression driven by a viral oncoprotein emerges as a crucial survival mechanism in Merkel cell carcinoma

doi: 10.1080/15548627.2025.2477385

Figure Lengend Snippet: MCPyV truncated LT induces paranuclear retention and stabilization of KIT. (A) Immunofluorescence detection of KIT (green) and MCPyV LT (CM2B4, red) or sT (CM8E6, red) in KIT-HEK293 cells transfected with MCPyV T-antigens or vector control ( n = 6). (B) Top : illustration of the expression constructs of LT339 and the VPS39-interaction defective mutant LT339 W209A . Bottom : Representative images showing the effect of LT339 and LT339 W209A mutant on localization of KIT (green). LT was detected by CM2B4 (red). ( n = 3) (C) Immunoblots showing the effect of the LT339 and LT339 W209A on KIT expression. The quantification of KIT level is shown below the immunoblots ( n = 9). (D) Top : immunoblot analysis of the effect LT339 and LT339 W209A on KIT protein stability in the presence of cycloheximide (CHX) up to 4 h. Bottom : quantification of KIT protein stability after normalization to 0 h time point ( n = 4). (E) Immunoblots showing KIT protein stability in KIT-HEK293 cells transfected with LT339, LT339 W209A or plasmid control (CTR) treated with KIT ligand (KITLG, 100 ng/mL) or solvent control (PBS) in the presence of CHX (100 μg/mL). (F) Quantification of KIT protein stability after normalization to 0 h time point ( n = 5). Solid lines represent PBS control (KITLG − ) and dotted lines represent KITLG treatment (KITLG + ). (A and B) Nuclei were stained by DAPI (blue). Scale bar: 10 μm. Numbers below the images refer to the proportion of cells with KIT paranuclear dot-like staining to the total number of cells analyzed. (C, D and F) Error bars represent mean ± SEM. * p < 0.05, *** p < 0.001, ns = not significant were calculated by one-way ANOVA with post-hoc Tukey’s test (C), two-way ANOVA (D) or two-way ANOVA with post-hoc Bonferroni’s test (F).

Article Snippet: For KITLG induced KIT degradation experiments, KIT-293 cells were transfected with LT339, LT339 W209A or CTR for 48 h, followed by addition of CHX (100 μg/mL) and KITLG (100 ng/mL; Proteintech Group, HZ-1024) or PBS.

Techniques: Immunofluorescence, Transfection, Plasmid Preparation, Control, Expressing, Construct, Mutagenesis, Western Blot, Solvent, Staining

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: