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: Cell Proliferation

Article Title: Expression and correlation of the Pi3k/Akt pathway and VEGF in oral submucous fibrosis

doi: 10.1111/cpr.13491

Figure Lengend Snippet: The effects of Pi3k/Akt signalling pathway on cell migration and proliferation. Relative protein expression after LY294002 and IGF‐1 treatment. (A) Images and (B) histograms of scratch tests on HUVECs treated with different concentrations of LY294002 for 0, 12 and 24 h. (C) Images and (D) histograms of scratch tests on HUVECs treated with different concentrations of IGF‐1 for 0, 12 and 24 h. (E‐H) Western Blot images of VEGF, Akt and p‐Akt expression at varying LY294002 concentrations. (J‐M) Western Blot images of VEGF, Akt and p‐Akt expression at varying IGF‐1 concentrations. (I, N) Histograms showing HUVEC viability after treatment with different concentrations of LY294002 and IGF‐1 for 24 h. Data are presented as mean ± standard deviation (SD) (n ≥ 3). Significance: * p < 0.05, ** p < 0.01, *** p < 0.001. (Scale bars = 200 μm).

Article Snippet: HUVECs (BNCC Co. Ltd.); fetal bovine serum (10%, HyClone); Penicillin–streptomycin solution (1%, HyClone); Optical microscope (DM2700 M; Leica Microsystems Technology Co. Ltd.); confocal microscopy (Olympus); whole cell lysis reagent (KeyGen Biotech); polyvinylidene fluoride (Beyotime); primary antibodies including anti‐Akt (1:1000, Abcam), anti‐p‐Akt (1:1000, Abcam), anti‐β‐actin (1:1000, CST, Boston), anti‐VEGF (1:1000, Abcam), anti‐Pi3k (1:1000, Abcam), anti‐p‐Pi3k (1:1000, Abcam), anti‐Col‐I (1:1000, CST, Boston); a gel and blot imaging system (Syngene); LY294002 (HY‐10108, MedChemExpress); IGF‐1 (HY‐P700478, MedChemExpress).

Techniques: Migration, Expressing, Western Blot, Standard Deviation

Journal: Cell Proliferation

Article Title: Expression and correlation of the Pi3k/Akt pathway and VEGF in oral submucous fibrosis

doi: 10.1111/cpr.13491

Figure Lengend Snippet: Pi3k/Akt pathway inhibitor (LY294002) and activator (IGF‐1) regulates VEGF and Akt expression at different concentrations. (A, B) VEGF fluorescence and expression after LY294002 treatment at varying doses. (C, D) Akt fluorescence and expression after LY294002 treatment at varying doses. (E, F) VEGF fluorescence and expression after IGF‐1 treatment at varying doses. (G, H) Akt fluorescence and expression after IGF‐1 treatment at varying doses.

Article Snippet: HUVECs (BNCC Co. Ltd.); fetal bovine serum (10%, HyClone); Penicillin–streptomycin solution (1%, HyClone); Optical microscope (DM2700 M; Leica Microsystems Technology Co. Ltd.); confocal microscopy (Olympus); whole cell lysis reagent (KeyGen Biotech); polyvinylidene fluoride (Beyotime); primary antibodies including anti‐Akt (1:1000, Abcam), anti‐p‐Akt (1:1000, Abcam), anti‐β‐actin (1:1000, CST, Boston), anti‐VEGF (1:1000, Abcam), anti‐Pi3k (1:1000, Abcam), anti‐p‐Pi3k (1:1000, Abcam), anti‐Col‐I (1:1000, CST, Boston); a gel and blot imaging system (Syngene); LY294002 (HY‐10108, MedChemExpress); IGF‐1 (HY‐P700478, MedChemExpress).

Techniques: Expressing, Fluorescence