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: Disease Markers

Article Title: Evaluation of Immune Infiltration Based on Image Plus Helps Predict the Prognosis of Stage III Gastric Cancer Patients with Significantly Different Outcomes in Northeastern China

doi: 10.1155/2022/2893336

Figure Lengend Snippet: Expression of CD4, CD8, CD20, CD56, CD68, CD117, and CD177 by immune cells infiltrated tumor tissues of GC patients. Representative images of immune markers staining in immune cells from GC samples are shown at ×200 (100 mm) original magnification. Positive expression of CD4 (a), CD8 (b), CD20 (c), CD56 (d), CD68 (e), CD117 (f), and CD177 (g) infiltrated in tumor tissues with a regular distribution. In order to show the distribution from immune cells more clearly, we used computerized imaging system Image-Pro Plus version 6.2 to highlight positive areas in different colors.

Article Snippet: After cleaned in distilled water, the paraffin sections were pretreated with citrate buffer, pH 6.0 (CD177) and EDTA Antigen Retrieval Solution, pH 8.0 (CD4, CD8, CD20, CD56, CD68, and CD117) for 3 min at 120°C in a pressure cooker, and endogenous peroxidase was inhibited with 3% H 2 O 2 in PBS for 10 min. Nonspecific actions in the sections were also blocked with goat serum (BOSTER, USA) for 1 h at room temperature.

Techniques: Expressing, Staining, Imaging

Journal: Disease Markers

Article Title: Evaluation of Immune Infiltration Based on Image Plus Helps Predict the Prognosis of Stage III Gastric Cancer Patients with Significantly Different Outcomes in Northeastern China

doi: 10.1155/2022/2893336

Figure Lengend Snippet: The schematic diagram of the distribution of immune cells. The blue points represent CD20 + B cells, the yellow points represent CD4 + T cells, the orange points represent CD117 + mast cells, the green points represent CD68 + macrophages, the pink points represent CD8 + T cells, the black points represent CD56 + NK cells, and the red points represent CD177 + neutrophils. Although not all immune cells are distributed according to this fixed law, this pattern can basically reflect the general characteristics of their distribution.

Article Snippet: After cleaned in distilled water, the paraffin sections were pretreated with citrate buffer, pH 6.0 (CD177) and EDTA Antigen Retrieval Solution, pH 8.0 (CD4, CD8, CD20, CD56, CD68, and CD117) for 3 min at 120°C in a pressure cooker, and endogenous peroxidase was inhibited with 3% H 2 O 2 in PBS for 10 min. Nonspecific actions in the sections were also blocked with goat serum (BOSTER, USA) for 1 h at room temperature.

Techniques:

Journal: Disease Markers

Article Title: Evaluation of Immune Infiltration Based on Image Plus Helps Predict the Prognosis of Stage III Gastric Cancer Patients with Significantly Different Outcomes in Northeastern China

doi: 10.1155/2022/2893336

Figure Lengend Snippet: Differences in immune marker positive area/total area between the two groups (survival time of patients in group A was less than 1 year and group B survival time was more than 5 years) by the rank sum test. (a) CD4 + T cells ( P < 0.001). (b) CD8 + T cells ( P < 0.001). (c) CD20 + B cells ( P < 0.001). (d) CD68 + macrophages ( P < 0.001). (e) CD117 + mast cells ( P < 0.001). (f) CD177 + neutrophils ( P < 0.001).

Article Snippet: After cleaned in distilled water, the paraffin sections were pretreated with citrate buffer, pH 6.0 (CD177) and EDTA Antigen Retrieval Solution, pH 8.0 (CD4, CD8, CD20, CD56, CD68, and CD117) for 3 min at 120°C in a pressure cooker, and endogenous peroxidase was inhibited with 3% H 2 O 2 in PBS for 10 min. Nonspecific actions in the sections were also blocked with goat serum (BOSTER, USA) for 1 h at room temperature.

Techniques: Marker