mt4 cells  (JCRB Cell Bank)

Journal: Nature Communications

Article Title: TSG-6+ cancer-associated fibroblasts modulate myeloid cell responses and impair anti-tumor response to immune checkpoint therapy in pancreatic cancer

doi: 10.1038/s41467-024-49189-x

Figure Lengend Snippet: a Schematic representation of the scRNAseq experimental design created with BioRender.com, released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license. 3 tumors in each group were pooled for internal control. b Representative Uniform Manifold Approximation Projection (UMAP) plot of sorted intratumoral CD45-negative cells. Each dot represents a cell. c UMAP plots indicating expression of genes depicting B16F10 tumor cells ( Pmel , Mlana ), mT4 tumor cells ( Krt18 , Krt19 ) and fibroblasts ( Col1a1 , Dcn ). d UMAP plots highlighting differences in fibroblast abundance between mT4 and B16F10 tumors (red circle). e UMAP plot depicting tnfaip6 (gene encoding TNF Stimulating Gene-6 (TSG-6)) expression in B16F10 and mT4 tumors (red circle). f Box-and-whisker plot representing TNFAIP6 RNA expression in TCGA datasets of melanoma (skcm, skin cutaneous melanoma) ( n = 480 patients) and pancreatic patient tumors (paad, pancreatic adenocarcinoma) ( n = 186 patients). Each dot represents a patient. Statistical significance was calculated using Student’s t test (two-tailed) and p value for the comparison has been indicated in the figure. Data are presented as mean values ± SD. The center of the plot represents mean of the group and the whiskers represent minimum- maximum values.

Article Snippet: mT4 pancreatic cell line was a generous gift from Dr. David A. Tuveson (Cold Spring Harbor Laboratory, NY). mT4 is an organoid cell line generated from mouse pancreata containing PDAC from the Kras LSL-G12D/+ ;Trp53 LSL-R172H/+ ;Pdx1–Cre mouse model under C57BL/6 background . mT4-LS cells were generated and generously gifted by Dr. Michael Curran (The University of Texas MD Anderson Cancer Center, Houston, TX).

Techniques: Control, Expressing, Whisker Assay, RNA Expression, Two Tailed Test, Comparison

Journal: Nature Communications

Article Title: TSG-6+ cancer-associated fibroblasts modulate myeloid cell responses and impair anti-tumor response to immune checkpoint therapy in pancreatic cancer

doi: 10.1038/s41467-024-49189-x

Figure Lengend Snippet: a Schematic representation of the scRNAseq experimental design created with BioRender.com, released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license. b Representative landscape in B16F10 and mT4 tumors. Three tumors in each group were pooled for internal control. All major immune cell subsets were identified. c Cluster frequency plot of each immune subset in B16F10 and mT4 tumors. The T cells are depicted in shades of green, B cells in purple, NK cells in gray, macrophages and monocytes in red, neutrophil in orange, and dendritic cells in blue. d Violin plot representing expression of marker genes used for characterization of immune subsets identified in ( b ). e UMAP plots depicting total macrophages in B16F10 tumors (red) and mT4 (blue) to highlight minimal overlap between subsets. Each dot represents a cell. f Distribution of the macrophages across the B16F10 and mT4 tumors depicted in ( e ). g Heatmap of functional markers for the individual macrophage subsets providing phenotypic information. Expression levels are scaled between minimum and maximum expression for each gene across all clusters. h GSEA results depicting differential pathways between mT4 and B16F10 macrophages.

Article Snippet: mT4 pancreatic cell line was a generous gift from Dr. David A. Tuveson (Cold Spring Harbor Laboratory, NY). mT4 is an organoid cell line generated from mouse pancreata containing PDAC from the Kras LSL-G12D/+ ;Trp53 LSL-R172H/+ ;Pdx1–Cre mouse model under C57BL/6 background . mT4-LS cells were generated and generously gifted by Dr. Michael Curran (The University of Texas MD Anderson Cancer Center, Houston, TX).

Techniques: Control, Expressing, Marker, Functional Assay

Journal: Nature Communications

Article Title: TSG-6+ cancer-associated fibroblasts modulate myeloid cell responses and impair anti-tumor response to immune checkpoint therapy in pancreatic cancer

doi: 10.1038/s41467-024-49189-x

Figure Lengend Snippet: a Expression of Cd44 across macrophages present in B16F10 and mT4 tumors. b Violin plot quantifying Cd44 expression in macrophages present in B16F10 and mT4 tumors. c Representative multi-immunofluorescence (mIF) image highlighting co-localization of CD68+ CD44+ CD163+ myeloid cells with TSG-6 (white arrow) in human pancreatic tissue FFPE samples. d Quantification of the mIF images using infiltration analysis technique. Red borders indicate TSG-6+ cells and areas from red to green indicate the increasing distance from the TSG-6+ cells (green being furthest). Percentage of CD68+ CD44+ cells that were at a distance of 0–20 μm (closest) from TSG6+ cells were quantified, and bar plotted ( n = 14 pancreatic tissues; patient characteristics provided in Supplementary Table ). e Quantification of number of CD68+ CD44+ cells that were at a distance of 0–20 μm (closest) from TSG-6+ SMA+ versus TSG-6+ SMA- cells ( n = 10 pancreatic tissues; patient characteristics provided in Supplementary Table ). Each symbol represents a patient. Statistical significance was calculated using Student’s t test (two-tailed). Data are presented as mean values ± SD and p values for each comparison has been indicated in the figure. Source data are provided as a file.

Article Snippet: mT4 pancreatic cell line was a generous gift from Dr. David A. Tuveson (Cold Spring Harbor Laboratory, NY). mT4 is an organoid cell line generated from mouse pancreata containing PDAC from the Kras LSL-G12D/+ ;Trp53 LSL-R172H/+ ;Pdx1–Cre mouse model under C57BL/6 background . mT4-LS cells were generated and generously gifted by Dr. Michael Curran (The University of Texas MD Anderson Cancer Center, Houston, TX).

Techniques: Expressing, Immunofluorescence, Two Tailed Test, Comparison