Journal: Clinical Cancer Research

Article Title: TAK1 Promotes an Immunosuppressive Tumor Microenvironment through Cancer-Associated Fibroblast Phenotypic Conversion in Pancreatic Ductal Adenocarcinoma

doi: 10.1158/1078-0432.CCR-24-1004

Figure Lengend Snippet: TAK1 activation is higher in CAFs in PDAC, associating with immunosuppressive markers, and TAK1 + CAFs are far from the α-SMA + phenotype CAFs. A, Box plots showing expression levels of TAK1 in PDAC tumors compared with normal samples in TCGA and GTEx data; n = 179 patients (tumor); n = 171 (normal). This analysis was conducted using one-way ANOVA. B, Representative co-IF image of TAK1 (red) and CK19 (green) in PDAC. C, Representative IHC staining of TAK1 in human PDAC tissues. Scale bar, 100 µm. D, Representative images of TAK1 (green) and PDPN (red) co-IF staining (left) and TAK1 (green) and FAP (red) co-IF staining (right) in PDAC. Scale bar, 100 µm. E, Western blots showing TAK1 expression in PDAC cell lines and CAFs. F, Correlation scatter plot with the Pearson coefficient ( R ) of the T-cell exhaustion signature ( LAG3 , PDCD1 , CTLA4 , HAVCR2 , and TIGHT ) vs MAP3K7 (log 2 RNA-seqV2, BE norm.) in the PDAC samples from TCGA database. G, Inverse correlation between TAK1 and α-SMA expression in CAF-infiltrating tumors in human PDAC slices. TAK1 (red) and α-SMA (green) expression in tumors in human PDAC slices were examined with IF staining; scale bar, 100 µm. Boxed areas ( a–c ) are magnified in adjacent panels, showing different levels of TAK1 and α-SMA expression in CAFs from patient sections. H, The CAFs were categorized into α-SMA + and α-SMA − groups, and the number of TAK1-positive cells was detected and quantified (as shown in G ). TAK1 positivity was analyzed and quantified in 50 randomly selected high-power fields (400×) from five patients with PDAC ( P < 0.01; unpaired Student t test ). I, Representative co-IF image of TAK1 (green), a-SMA (green), and CK19 (red). The images on the right are enlarged views. The white lines represent the tumor edge and CAF edge. Arrows indicate the vertical distance between the two white lines. TAK1 + cells infiltrated into the stroma, where α-SMA + CAFs were near the cancer gland. Counterstaining with DAPI (blue). Bar, 100 μm (bottom). J, Quantification calculation of the vertical distance between TAK1 + CAFs with CK19 and between α-SMA + CAFs with CK19 in the stroma. Results show mean ± SD of five human tissues. ****, P < 0.0001, unpaired Student t test. DAPI, 4′,6-diamidino-2-phenylindole; FAP, fibroblast activation protein; GTEx, Genotype-Tissue Expression; PAAD, pancreatic adenocarcinoma; PDPN, podoplanin.

Article Snippet: Human primary active CAFs (CAF1, CAF2, and CAF3) were derived from fresh pancreatic cancer surgical specimens at Kyushu University, following the method described by Bachem and colleagues ( ) and Ikenaga and colleagues ( ) These isolated cells exhibited fibroblast-like morphology and tested negative for cytokeratin 19 (CK19), an epithelial cell marker ( ).

Techniques: Activation Assay, Expressing, Immunohistochemistry, Staining, Western Blot

Journal: Clinical Cancer Research

Article Title: TAK1 Promotes an Immunosuppressive Tumor Microenvironment through Cancer-Associated Fibroblast Phenotypic Conversion in Pancreatic Ductal Adenocarcinoma

doi: 10.1158/1078-0432.CCR-24-1004

Figure Lengend Snippet: Interference with TAK1 transforms CAFs into myCAFs, inhibits MAPK and NF-κB pathways, reduces the secretion of inflammatory factors, and inhibits related oncogenes. A, WB indicated protein expression levels of CAFs after treatment with OXO at 100, 200, and 300 nmol/L. The quantitative analysis of proteins was measured by their density. B, The data are expressed as mean ± SD. Statistical significance was determined using an unpaired two-tailed t test. *, P < 0.05; **, P < 0.01; ***, P < 0.001. C, TAK1 knockdown of CAFs with siRNA. The effects of α-SMA protein and IL6 protein levels are shown through IF; scale bar, 100 μm. D, Western blot analysis of the levels of TAK1, p-TAK1 (phosphorylation of TAK1), P38, p-P38 (phosphorylation of P38), NF-κB, and p-NFκB in CAFs by TAK1 knockdown. E, Primary human CAFs obtained from two distinct patients with PDAC (CAF1 or CAF2) underwent two stable TAK1 knockdown and Western blot analyses of oncoprotein expression in CAFs. F, A human cytokine array of CAFs was treated with OXO at 300 nmol/L. The most significant gene alterations are displayed in the chart. DAPI, 4′,6-diamidino-2-phenylindole; OPG, osteoprotegerin.

Article Snippet: Human primary active CAFs (CAF1, CAF2, and CAF3) were derived from fresh pancreatic cancer surgical specimens at Kyushu University, following the method described by Bachem and colleagues ( ) and Ikenaga and colleagues ( ) These isolated cells exhibited fibroblast-like morphology and tested negative for cytokeratin 19 (CK19), an epithelial cell marker ( ).

Techniques: Expressing, Two Tailed Test, Knockdown, Western Blot, Phospho-proteomics

Journal: Micromachines

Article Title: Fragmentation Characteristics of Bubbles in a Throttling Hole Pipe

doi: 10.3390/mi15081025

Figure Lengend Snippet: Results of Physical Model Testing and Validation.

Article Snippet: The turbulence model used in the COMSOL commercial software is based on the Reynolds-averaged equations, with commonly used types including the standard k-ε model, the RNG k-ε model, and the Reynolds stress model [ , ].

Techniques: Biomarker Discovery