Journal: Cell Communication and Signaling : CCS

Article Title: Aldolase a coordinates macropinocytic nutrient scavenging and lysosomal degradation in lung cancer by interacting with V-ATPase

doi: 10.1186/s12964-025-02591-4

Figure Lengend Snippet: Targeting ALDOA impairs lysosomal proteolysis and tumor growth in vivo. A Immunoblot analysis confirming the effect of Aldolase A (ALDOA)-targeting sgRNA on protein levels of ALDOA in A549 cells. B Effects of bovine serum albumin (BSA) supplementation on proliferation of sgALDOA cells under leucine-replete and leucine-deprived conditions. C , D Representative gross images ( C ) and growth curves ( D ) of xenografted tumors derived from A549 control or sgALDOA cells, treated with or without the macropinocytosis inhibitor 5-(N-Ethyl-N-isopropyl)amiloride (EIPA) ( N = 6 per group). E Representative immunofluorescence images of Lysosome (red) and ALDOA (green) in tumor tissues from xenografted A549 models. Arrows indicate colocalized puncta. F Representative images of DQ-BSA fluorescence (left panel) and quantification of DQ-BSA fluorescence intensity (right panel) in tumor tissue formed after subcutaneous injection of Ctrl or sgALDOA A549 cells, with or without EIPA treatment. G Representative histological (H&E) and immunohistochemical images of Ki67 and p-S6 staining (left panel), and quantification of Ki67 and p-S6 expression scores (right panel), in xenograft tumor sections. H Representative immunofluorescence images of ALDOA (green) and α-smooth muscle actin (α-SMA, red) in xenograft tumor sections (left panel), and quantification of ALDOA fluorescence in α-SMA–negative cells (right panel). All experimental data were verified in at least six independent experiments. Scale bar, 10 μm. Data are presented as the mean ± SEM. ns., not significant, ** p < 0.01, and *** p < 0.001

Article Snippet: For tumor tissue staining, deparaffinized samples were incubated with anti-ALDOA (Proteintech) and anti-LAMP1 (Santa Cruz) or anti-ALDOA (Santa Cruz) and anti-α-smooth muscle actin (α-SMA) (Cell Signaling).

Techniques: In Vivo, Western Blot, Derivative Assay, Control, Immunofluorescence, Fluorescence, Injection, Immunohistochemical staining, Staining, Expressing

Journal: Annals of Gastroenterological Surgery

Article Title: Intratumoral Fusobacterium nucleatum Drives Cancer‐Associated Fibroblasts Enrichment and Immune Exclusion in Esophageal Squamous Cell Carcinoma

doi: 10.1002/ags3.70116

Figure Lengend Snippet: Histopathological assessment of CAFs in ESCC. (a) Representative FISH images of ESCC tissues showing signals for F. nucleatum (red; FUS664), all bacteria (green; EUB338), and nuclei (blue; DAPI). Positive F. nucleatum signals were observed in cases that were identified as F. nucleatum –positive by qPCR. All images were acquired at ×200 magnification. (b) Representative FISH images of ESCC tissues obtained by laser scanning confocal microscopy, demonstrating intracellular signals of Fusobacterium nucleatum (red; FUS664) within tumor cells. Nuclei were counterstained with DAPI (blue). Images were acquired at ×400 magnification. (c) Representative FISH images of ESCC tissues showing signals for F. nucleatum (red; FUS664), all bacteria (green; EUB338), and nuclei (blue; DAPI). F. nucleatum signals were observed within tumor cells located in regions enriched with α‐SMA–positive CAFs. The boxed areas indicate regions of interest, imaged at ×200 magnification. ** p < 0.01.

Article Snippet: We used monoclonal mouse anti α‐SMA antibody (1:250; #56856; Cell Signal Technology) and RelA (1:800; #8242; Cell Signal Technology) as the primary antibody, following the manufacturer's protocols.

Techniques: Bacteria, Confocal Microscopy

Journal: Annals of Gastroenterological Surgery

Article Title: Intratumoral Fusobacterium nucleatum Drives Cancer‐Associated Fibroblasts Enrichment and Immune Exclusion in Esophageal Squamous Cell Carcinoma

doi: 10.1002/ags3.70116

Figure Lengend Snippet: Immunohistochemical analysis of NF‐κB activation and its association with F. nucleatum and CAFs in ESCC. (a) The proportion of NF‐κB–positive tumors was significantly higher in F. nucleatum –positive cases than in F. nucleatum –negative cases. (b) Representative immunohistochemical staining on adjacent serial sections of ESCC tissues showing stromal α‐SMA expression and nuclear RelA localization in tumor cells. These signals were observed in close proximity. Images were acquired at ×200 magnification. (c) Dual positivity for stromal α‐SMA and NF‐κB–positive in tumor cells was significantly enriched in F. nucleatum –positive tumors compared to F. nucleatum –negative tumors. (d) Summary of the results. F. nucleatum contributes to the progression of ESCC by inducing NF‐κB–mediated inflammatory signaling in tumor cells and promoting the activation of CAFs. ** p < 0.01.

Article Snippet: We used monoclonal mouse anti α‐SMA antibody (1:250; #56856; Cell Signal Technology) and RelA (1:800; #8242; Cell Signal Technology) as the primary antibody, following the manufacturer's protocols.

Techniques: Immunohistochemical staining, Activation Assay, Staining, Expressing

Journal: Journal of Extracellular Vesicles

Article Title: Stromal cell‐derived small extracellular vesicles enhance radioresistance of prostate cancer cells via interleukin‐8‐induced autophagy

doi: 10.1002/jev2.12342

Figure Lengend Snippet: sEVs derived from CAFs enhance the radioresistance of PCa cells. (a) CAFs were isolated from prostate primary tumour tissues, and the specific phenotype was examined under microscopy. (b,c) The isolated CAFs were further verified by flow cytometry using antibodies against fibroblastic markers (α‐SMA and FAP) and PCa markers (PSA and AR). (d) The levels of IL‐8 in CAF‐derived sEVs and stromal cell‐derived sEVs were measured using its ELISA kit ( n = 3). (e) sEVs were isolated from CAF cultural supernatant and characterized by immunoblotting. (f) CAF‐derived sEVs were cocultured with 22Rv1 cells before IR treatment. Cytotoxicity was quantified using a CCK‐8 assay ( n = 3). (g‐j) Furthermore, the effect of CAF‐sEVs on PCa cell survival was confirmed using colony survival assay ( n = 3). **( p < 0.01) and ***( p < 0.001) show the significances between different groups as indicated; ns shows no significance.

Article Snippet: The cultured CAFs were harvested and characterized using FACSCalibur Flow Cytometry (BD Sciences, San Jose, CA, USA) with primary antibodies against FAP (#66562, CST, Danvers, MA, USA) and α‐SMA (#69319, CST) with secondary antibodies conjugated with red or green fluorescent dye (#S0014, #S0018, Affinity).

Techniques: Derivative Assay, Isolation, Microscopy, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Western Blot, CCK-8 Assay, Clonogenic Cell Survival Assay