Journal: Advanced Science

Article Title: The Characteristics of Tumor Microenvironment by the Single Cell Profile in Endometrial Cancer of no Specific Molecular Profile (NSMP) Subtype

doi: 10.1002/advs.78332

Figure Lengend Snippet: SPP1+ macrophages promote T cell stress and maintain epithelial cell stemness. (A, B) Western blot and RT‐qPCR results of SPP1 expression in macrophages with SPP1 knockdown. (C, D) Western blot and RT‐qPCR results of SPP1 expression in macrophages with overexpression of SPP1. (E) Western blot analysis of the protein expression of HSP70, LAG3, and PD‐1 in Jurkat cells after co‐culture with control and SPP1 knockdown macrophages. (F) qRT‐PCR analysis of the transcriptional level of HSPA1A and HSPA1B in Jurkat cells after co‐culture with control and SPP1 knockdown macrophages. (G) Western blot analysis of the protein expression of HSP70, LAG3, and PD‐1 in Jurkat cells after co‐culture with control, SPP1 overexpression, and SPP1 overexpression plus anti‐CD44 macrophages. (H) qRT‐PCR analysis of the transcriptional level of HSPA1A and HSPA1B in Jurkat cells after co‐culture with control, SPP1 overexpression, and SPP1 overexpression plus anti‐CD44 macrophages. (I) Western blot analysis of the protein expression of CD44 and CD133 in Ishikawa cells after co‐culture with control and SPP1 knockdown macrophages. (J) Western blot analysis of the protein expression of CD44 and CD133 in Ishikawa cells after co‐culture with control, SPP1 overexpression, and SPP1 overexpression plus anti‐CD44 macrophages. (K) Sphere formation assay was used to detect the stemness ability of Ishikawa cells after co‐culture with control and SPP1 knockdown macrophages. (L) A sphere formation assay was used to detect the stemness ability of Ishikawa cells after co‐culture with control, SPP1 overexpression, and SPP1 overexpression plus anti‐CD44 macrophages. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns: not significant.

Article Snippet: The primary antibodies used in this study were as follows: CD44 (15675‐1‐AP, Proteintech, 1:20000), SPP1 (22952‐1‐AP, Proteintech, 1:1000), CD133 (18470‐1‐AP, Proteintech, 1:5000), HSP70 (80735‐1‐RR, Proteintech, 1:10000), PD‐1 (66220‐1‐Ig, Proteintech, 1:20000), LAG‐3 (16616‐1‐AP, Proteintech, 1:1000) and β‐actin (66009‐1‐Ig, Proteintech, 1:20000).

Techniques: Western Blot, Quantitative RT-PCR, Expressing, Knockdown, Over Expression, Analysis, Co-culture Assay, Control, Tube Formation Assay

Journal: Translational Oncology

Article Title: MSRB3 promotes gastric cancer progression and immunotherapy resistance by stabilizing GPX4 to inhibit ferroptosis

doi: 10.1016/j.tranon.2026.103081

Figure Lengend Snippet: MSRB3 deficiency enhances anti-PD-1 efficacy in a ferroptosis-dependent manner and augments CD8⁺ T cell infiltration in vivo . (A) Knockdown efficiency of MSRB3 in MFC cells was confirmed by western blotting. (B–D) Representative tumor images (B), tumor growth curves (C), and tumor weights (D) of xenograft models treated with anti–PD-1 antibody and/or Fer-1. MSRB3 knockdown synergized with PD-1 blockade to suppress tumor growth, which was reversed by Fer-1 treatment. (E, F) Representative Ki-67 staining and quantification showing markedly reduced tumor proliferation in the combination treatment group. Scale bar, 50 μm. (G, H) Representative immunofluorescence images and quantification of CD8⁺ T-cell infiltration in xenograft tumors. Scale bar, 50 μm. Data are presented as mean ± SD (n = 5 mice per group). * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Anti-PD-1 antibody (RMP1–14) was purchased from BioXCell (West Lebanon, NH, USA).

Techniques: In Vivo, Knockdown, Western Blot, Xenograft Assay, Staining, Immunofluorescence

Journal: Advanced Science

Article Title: Oral Astilbin Nanogels Modulate Senescence‐Associated CD4 + T Cells and Promote T reg Induction in Aged Mice with Colitis

doi: 10.1002/advs.78123

Figure Lengend Snippet: Ast attenuates senescence‐associated features and restores the immunoregulatory function of CD4 + T cells. (A) Correlation between PDCD1 expression, which encodes PD‐1, and the age at diagnosis in patients with colitis ( n = 106). (B) Comparison of PDCD1 expression levels between young (< 60 years old) and aged (≥ 60 years old) colitis patients. (C) Experimental flowchart for DSS‐induced colitis in young and aged mice. (D) Curve chart analysis of DAI ( n = 6). (E) Representative images of colons and corresponding quantification of colon length ( n = 6). (F) Quantitative analysis of CD4 + PD‐1 + T cells and CD4 + SA‐β‐gal + T cells in colonic lamina propria cells and representative FC plots ( n = 6). (G) In vitro effects of different concentrations of Ast on PD‐1 expression in colonic CD4 + T cells isolated from young and aged mice ( n = 5). (H) In vitro proliferation of CD4 + T cells isolated from aged mice following Ast treatment ( n = 3). (I) In vitro differentiation of CD4 + T cells isolated from aged mice into T regs following Ast treatment ( n = 3). Data are presented as mean ± SD and analyzed by two‐tailed unpaired Student's t test with Welch's correction (B), two‐tailed unpaired Student's t test (H, I), Two‐way ANOVA with Tukey's post‐test (D), or One‐way ANOVA with Tukey's post‐test (E–G). ns represents not significant, * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: Purified blocking anti‐mouse PD‐1 (Clone RMP1‐14), agonistic anti‐mouse PD‐1 (Clone RMP1‐30), anti‐mouse CD3 (Clone 17A2), and anti‐mouse CD28 (Clone 37.51) were purchased from BioXcell (USA).

Techniques: Expressing, Biomarker Discovery, Comparison, Analysis, In Vitro, Isolation, Two Tailed Test

Journal: Advanced Science

Article Title: Oral Astilbin Nanogels Modulate Senescence‐Associated CD4 + T Cells and Promote T reg Induction in Aged Mice with Colitis

doi: 10.1002/advs.78123

Figure Lengend Snippet: High‐dose free Ast alleviates colitis in young and aged mice by attenuating senescence‐associated immune dysfunction. (A) Experimental flowchart of young and aged colitis mice treated with free Ast. (B) Curve chart variations of murine body weight across time, normalized to the day 0 body weight percentage ( n = 6). (C) Curve chart analysis of DAI ( n = 6). (D) Representative images of colons and corresponding quantification of colon length in young and aged mice ( n = 6). Representative FC plots and quantitative analysis of (E) CD4 + PD‐1 + T cells, (F) CD4 + SA‐β‐gal + T cells, and (G) CD25 + Foxp3 + CD4 T cells in colonic lamina propria cells ( n = 6). (H) Heatmap of differentially expressed genes in colon samples ( n = 6). GSEA of the (I) TGF‐beta signaling pathway and (J) AGE‐RAGE signaling pathway. Data are presented as mean ± SD and analyzed by One‐way ANOVA with Tukey's post‐test (D‐G) or Two‐way ANOVA with Tukey's post‐test (B, C). ns represents not significant, * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: Purified blocking anti‐mouse PD‐1 (Clone RMP1‐14), agonistic anti‐mouse PD‐1 (Clone RMP1‐30), anti‐mouse CD3 (Clone 17A2), and anti‐mouse CD28 (Clone 37.51) were purchased from BioXcell (USA).

Techniques: Analysis

Journal: Advanced Science

Article Title: Oral Astilbin Nanogels Modulate Senescence‐Associated CD4 + T Cells and Promote T reg Induction in Aged Mice with Colitis

doi: 10.1002/advs.78123

Figure Lengend Snippet: Mechanistic validation of Ast‐mediated regulation of senescence‐associated CD4 + T cells in aged colitis. (A) Experimental flowchart of NG treatment in aged colitis mice ( n = 6). (B) Representative fecal samples following occult blood testing. (C) Curve chart analysis of DAI ( n = 6). (D) Representative images of colons and corresponding quantification of colon length ( n = 6). (E) Quantification of PD‐1 + CD4 T cells in colonic lamina propria cells ( n = 6). (F) Representative FC plots and quantitative analysis of CD4 + CD27 − CD28 − T cells in colonic lamina propria cells ( n = 6). (G) Schematic illustration of the isolation and sorting strategy for senescence‐associated (CD28 − CD27 − ) and non‐senescence‐associated (remaining) CD4 + T cells. Created with BioRender. (H) MFI of PD‐1 expression on senescence‐associated and non‐senescence‐associated CD4 + T cells after treatment with PBS or Ast for 2 days ( n = 3). (I) Cell viability of senescence‐associated and non‐senescence‐associated CD4 + T cells ( n = 3). (J) Foxp3 + T reg differentiation capacity of senescence‐associated and non‐senescence‐associated CD4 + T cells in the presence of PBS or Ast for 2 days ( n = 3). (K) Experimental groups for therapeutic evaluation in aged colitis mice. (L) Curve chart analysis of DAI ( n = 6). (M) Quantification of colon length ( n = 6). Quantitative analysis of (N) CD4 + PD‐1 + T cells and (O) CD4 + CD27 − CD28 − T cells in colonic lamina propria cells ( n = 6). (P) Schematic illustration of CD4 + T cells cultured in the presence of plate‐bound agonistic aPD‐1 or Ast. Created with BioRender. (Q) Quantitative analysis of CD4 + CD27 − CD28 − T cells following treatment with plate‐bound agonistic aPD‐1 or Ast ( n = 3). Data are presented as mean ± SD and analyzed by Two‐way ANOVA with Tukey's post‐test (C, L) or One‐way ANOVA with Tukey's post‐test (D‐F, H‐J, M‐O, Q). ns represents not significant, * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: Purified blocking anti‐mouse PD‐1 (Clone RMP1‐14), agonistic anti‐mouse PD‐1 (Clone RMP1‐30), anti‐mouse CD3 (Clone 17A2), and anti‐mouse CD28 (Clone 37.51) were purchased from BioXcell (USA).

Techniques: Biomarker Discovery, Fecal/Stool Samples, Analysis, Isolation, Expressing, Cell Culture, FACS