cell signaling technology 68790  (Cell Signaling Technology Inc)

Journal: bioRxiv

Article Title: Somatic TYK2 activating mutations in tumor-infiltrating T cells promote anti-cancer immunity

doi: 10.1101/2025.10.13.682217

Figure Lengend Snippet: NanoSeq of TILs identified somatic mutations in TYK2 . a. A schematic of somatic genomic sequencing of patient derived TILs. b. Distribution of tumor patients types. c. Classification of TYK2 mutation types in TILs. d. Lollipop plots of TYK2 mutations in tumor-infiltrating CD4⁺ T cells and tumor-infiltrating CD8⁺ T cells.

Article Snippet: The membranes were blocked with 5%(w/v) skim milk (BD Bioscience, USA) for 1 h at room temperature, and then incubated with the following primary antibodies overnight at 4 °C: TYK2 (Proteintech #16412-1-AP, 1:1000), TYK2 (Cell Signaling Technology #35615S, 1:1000), Phospho-TYK2 (Cell Signaling Technology, 68790S, 1:1000), STAT1 (Cell Signaling Technology, 14994S, 1:1000), Phospho-STAT1 (Cell Signaling Technology, 9167S, 1:1000), STAT3 (Cell Signaling Technology, 9139T, 1:1000), Phospho-STAT3 (Cell Signaling Technology, 9145S, 1:1000) and β-actin (Cell Signaling #4970, 1:1000).

Techniques: Genomic Sequencing, Derivative Assay, Mutagenesis

Journal: bioRxiv

Article Title: Somatic TYK2 activating mutations in tumor-infiltrating T cells promote anti-cancer immunity

doi: 10.1101/2025.10.13.682217

Figure Lengend Snippet: Lollipop plots of TYK2 mutations in TILs. a. Locations of TYK2 mutations in tumor-infiltrating B cells and tumor-infiltrating NK cells.

Article Snippet: The membranes were blocked with 5%(w/v) skim milk (BD Bioscience, USA) for 1 h at room temperature, and then incubated with the following primary antibodies overnight at 4 °C: TYK2 (Proteintech #16412-1-AP, 1:1000), TYK2 (Cell Signaling Technology #35615S, 1:1000), Phospho-TYK2 (Cell Signaling Technology, 68790S, 1:1000), STAT1 (Cell Signaling Technology, 14994S, 1:1000), Phospho-STAT1 (Cell Signaling Technology, 9167S, 1:1000), STAT3 (Cell Signaling Technology, 9139T, 1:1000), Phospho-STAT3 (Cell Signaling Technology, 9145S, 1:1000) and β-actin (Cell Signaling #4970, 1:1000).

Techniques:

Journal: bioRxiv

Article Title: Somatic TYK2 activating mutations in tumor-infiltrating T cells promote anti-cancer immunity

doi: 10.1101/2025.10.13.682217

Figure Lengend Snippet: TYK2 D810V expression enhances T cells effector function a. Western blot of Jurkat cells overexpressing 15 TYK2 missense mutations. Representative blot from two independent experiments. b. IFNγ and IL8 levels measured in primary T cells expressing empty vector, TYK2 WT , or TYK2 D810V with or without IL12 ( n= 3 replicates per group). c. Representative images of A549 tumor co-cultured with primary T cells overexpressed empty vector, TYK2 WT , or TYK2 D810V , scale bars = 1000 μm ( n= 3 replicates per group). d. Relative tumor area of A549 tumor co-cultured with primary T cells overexpressed empty vector, TYK2 WT , or TYK2 D810V ( n =3 replicates per group). e. IFNγ levels measured in A549 tumor cells co-cultured with primary T cells expressing empty vector, TYK2 WT , or TYK2 D810V ( n =3 replicates per group). f. Volcano plot showing upregulated genes in Jurkat T cells overexpressing TYK2 D810V compared to TYK2 WT ( n= 3 replicates per group). g. Enriched gene signatures associated with up- or down-regulated genes associated with TYK2 D810V compared to TYK2 WT . h. GSEA showing upregulation of Interferon-gamma response, Interferon-alpha response, Interferon-gamma response and IL6-JAK-STAT3 signaling genes in TYK2 D810V overexpressing Juakat T cells.

Article Snippet: The membranes were blocked with 5%(w/v) skim milk (BD Bioscience, USA) for 1 h at room temperature, and then incubated with the following primary antibodies overnight at 4 °C: TYK2 (Proteintech #16412-1-AP, 1:1000), TYK2 (Cell Signaling Technology #35615S, 1:1000), Phospho-TYK2 (Cell Signaling Technology, 68790S, 1:1000), STAT1 (Cell Signaling Technology, 14994S, 1:1000), Phospho-STAT1 (Cell Signaling Technology, 9167S, 1:1000), STAT3 (Cell Signaling Technology, 9139T, 1:1000), Phospho-STAT3 (Cell Signaling Technology, 9145S, 1:1000) and β-actin (Cell Signaling #4970, 1:1000).

Techniques: Expressing, Western Blot, Plasmid Preparation, Cell Culture

Journal: bioRxiv

Article Title: Somatic TYK2 activating mutations in tumor-infiltrating T cells promote anti-cancer immunity

doi: 10.1101/2025.10.13.682217

Figure Lengend Snippet: Functional characterization of TYK2 D810V mutation and single-cell transcriptomic profiling of TYK2-deficient primary T cells. a . STAT3 reporter activity in TYK2-knockout 293T cells expressing empty vector, TYK2 WT or TYK2 D810V . b. A schematic of A549 tumor and T cell co-culture experiment. c. Volcano plot showing genes downregulated in TYK2-knockout primary human CD4⁺ T cells. d. Volcano plot showing genes downregulated in TYK2-knockout primary human CD8⁺ T cells. e. GSEA enrichment plot showing top enriched Hallmark gene sets ranked by NES in TYK2-knockout primary CD4⁺ T cells. f. GSEA enrichment plot showing top enriched Hallmark gene sets ranked by NES in TYK2-knockout primary CD8⁺ T cells.

Article Snippet: The membranes were blocked with 5%(w/v) skim milk (BD Bioscience, USA) for 1 h at room temperature, and then incubated with the following primary antibodies overnight at 4 °C: TYK2 (Proteintech #16412-1-AP, 1:1000), TYK2 (Cell Signaling Technology #35615S, 1:1000), Phospho-TYK2 (Cell Signaling Technology, 68790S, 1:1000), STAT1 (Cell Signaling Technology, 14994S, 1:1000), Phospho-STAT1 (Cell Signaling Technology, 9167S, 1:1000), STAT3 (Cell Signaling Technology, 9139T, 1:1000), Phospho-STAT3 (Cell Signaling Technology, 9145S, 1:1000) and β-actin (Cell Signaling #4970, 1:1000).

Techniques: Functional Assay, Mutagenesis, Activity Assay, Knock-Out, Expressing, Plasmid Preparation, Co-Culture Assay

Journal: bioRxiv

Article Title: Somatic TYK2 activating mutations in tumor-infiltrating T cells promote anti-cancer immunity

doi: 10.1101/2025.10.13.682217

Figure Lengend Snippet: Design of Tyk2 D807V mutant mice. a. Genomic sequences for human TYK2 D810V and mouse Tyk2 D807V . b. Single-stranded DNA (ssDNA) and sgRNA used with CAS9 to generate the Tyk2 D807V mouse. c. Mouse TYK2 genomic sequences before and after CRISPR editing. d. Sanger sequencing for het Tyk2 D807V point mutation mice.

Article Snippet: The membranes were blocked with 5%(w/v) skim milk (BD Bioscience, USA) for 1 h at room temperature, and then incubated with the following primary antibodies overnight at 4 °C: TYK2 (Proteintech #16412-1-AP, 1:1000), TYK2 (Cell Signaling Technology #35615S, 1:1000), Phospho-TYK2 (Cell Signaling Technology, 68790S, 1:1000), STAT1 (Cell Signaling Technology, 14994S, 1:1000), Phospho-STAT1 (Cell Signaling Technology, 9167S, 1:1000), STAT3 (Cell Signaling Technology, 9139T, 1:1000), Phospho-STAT3 (Cell Signaling Technology, 9145S, 1:1000) and β-actin (Cell Signaling #4970, 1:1000).

Techniques: Mutagenesis, Genomic Sequencing, CRISPR, Sequencing

Journal: bioRxiv

Article Title: Somatic TYK2 activating mutations in tumor-infiltrating T cells promote anti-cancer immunity

doi: 10.1101/2025.10.13.682217

Figure Lengend Snippet: TYK2 D807V mice do not exhibit signs of autoimmunity or premalignancy. a. Representative appearance of 2- and 12-month-old het or homo Tyk2 D807V and Tyk2 WT mice. b. Body weights of the above mice. Males: WT ( n =8); het ( n =12); homo ( n =7). Females: WT ( n =6); het ( n =11); homo ( n =6). c. Serum levels of ALT and AST (U/L) of the above mice (WT ( n = 6); het ( n = 6); homo ( n = 6). mean ± s.e.m. is depicted. d. Representative H&E-stained liver sections from above mice (scale bars = 250 μm). e. Representative H&E-stained splenic sections from above mice (scale bars = 250 μm). f. Complete blood counts from 12-month-old mice. g. Representative spleens from above mice (scale bars = 10 mm). h. Representative western blot of liver tissues from 2-month old mice. For the data in this figure, each dot is one mouse. Error bars show mean ± s.e.m. P values were determined by one-way ANOVA followed by Tukey’s multiple comparisons test. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001; ns, not significant.

Article Snippet: The membranes were blocked with 5%(w/v) skim milk (BD Bioscience, USA) for 1 h at room temperature, and then incubated with the following primary antibodies overnight at 4 °C: TYK2 (Proteintech #16412-1-AP, 1:1000), TYK2 (Cell Signaling Technology #35615S, 1:1000), Phospho-TYK2 (Cell Signaling Technology, 68790S, 1:1000), STAT1 (Cell Signaling Technology, 14994S, 1:1000), Phospho-STAT1 (Cell Signaling Technology, 9167S, 1:1000), STAT3 (Cell Signaling Technology, 9139T, 1:1000), Phospho-STAT3 (Cell Signaling Technology, 9145S, 1:1000) and β-actin (Cell Signaling #4970, 1:1000).

Techniques: Staining, Western Blot

Journal: bioRxiv

Article Title: Somatic TYK2 activating mutations in tumor-infiltrating T cells promote anti-cancer immunity

doi: 10.1101/2025.10.13.682217

Figure Lengend Snippet: Tyk2 D807V mutant mice appear healthy with no obvious signs of disease. a. H&E of kidney sections of 2- and 12-month-old het or homo Tyk2 D807V mice and WT littermates (scale bars = 250 μm). b. Spleen-to-body weight ratios (n = 6 for each genotype). c. Complete blood counts of 2-month-old het or homo Tyk2 D807V mice and WT littermates (n = 6 for each genotype). For data in this figure, mean ± s.e.m. is depicted. P values were determined by one-way ANOVA followed by Tukey’s multiple comparisons test. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001; ns, not significant.

Article Snippet: The membranes were blocked with 5%(w/v) skim milk (BD Bioscience, USA) for 1 h at room temperature, and then incubated with the following primary antibodies overnight at 4 °C: TYK2 (Proteintech #16412-1-AP, 1:1000), TYK2 (Cell Signaling Technology #35615S, 1:1000), Phospho-TYK2 (Cell Signaling Technology, 68790S, 1:1000), STAT1 (Cell Signaling Technology, 14994S, 1:1000), Phospho-STAT1 (Cell Signaling Technology, 9167S, 1:1000), STAT3 (Cell Signaling Technology, 9139T, 1:1000), Phospho-STAT3 (Cell Signaling Technology, 9145S, 1:1000) and β-actin (Cell Signaling #4970, 1:1000).

Techniques: Mutagenesis

Journal: bioRxiv

Article Title: Somatic TYK2 activating mutations in tumor-infiltrating T cells promote anti-cancer immunity

doi: 10.1101/2025.10.13.682217

Figure Lengend Snippet: TYK2 D807V mice show enhanced anti-tumor effects. a. Syngeneic MC38 growth in male Tyk2 WT , het, or homo Tyk2 D807V mice. The tumor number is shown as the “n” in the figure and two tumors per mouse were implanted. P values were determined by two-way ANOVA. b. Tumor weights from panel a . P values were determined by one-way ANOVA followed by Tukey’s multiple comparisons test. c. Representative tumors from each group in panel a were ranked by size in descending order and the top six are shown. d. Syngeneic MC38 growth in female het Tyk2 D807V mice and Tyk2 WT littermates. The tumor number is shown as the “n” in the figure and two tumors per mouse were implanted. P values were determined by two-way ANOVA. e. Quantification of tumor-infiltrating CD45⁺ cells analyzed by flow cytometry. Each data point indicates an individual tumor (WT ( n = 8); Het ( n = 8)). P values were determined using two-tailed unpaired t -tests. f. Quantification of CD4⁺ and CD8⁺ T cell subsets from tumors analyzed by flow cytometry. Each data point indicates an individual tumor (WT ( n = 8); Het ( n = 8)). P values were determined using two-tailed unpaired t -tests. g. Syngeneic MC38 growth +/- anti-PD-1 antibodies. The tumor number is shown as the “n” in the figure and two tumors per mouse were implanted. P values were determined by two-way ANOVA. h. Tumor weight collected from panel g . P values were determined by one-way ANOVA followed by Tukey’s multiple comparisons test. i. Representative tumors from each group in panel g were ranked by size in descending order and the top six are shown. j. Syngeneic RIL175 growth, +/- anti-PD-1 antibodies. The tumor number is shown as the “n” in the figure and two tumors per mouse were implanted. P values were determined by two-way ANOVA. k. Tumor weight collected from panel j . P values were determined by one-way ANOVA followed by Tukey’s multiple comparisons test. l. Representative tumors from each group in panel j were ranked by size in descending order and the top six are shown. m. Syngeneic B16 growth, +/- anti-PD-1 antibodies. The tumor number is shown as the “n” in the figure and two tumors per mouse were implanted. P values were determined by two-way ANOVA. n. Tumor weight collected from panel m . P values were determined by one-way ANOVA followed by Tukey’s multiple comparisons test. o. Representative tumors from each group in panel m were ranked by size in descending order and the top six are shown. p. Representative livers of Tyk2 D807V het and WT littermates with shp53/ NRAS G12V tumors, +/- anti-PD-1 antibodies (scale bar = 10 mm). q. Liver-to-body weight ratios from panel p . Each data point is one liver and P values were determined by one-way ANOVA followed by Tukey’s multiple comparisons test. r. Representative livers of Tyk2 D807V het and WT littermates with CTNNB1 / MYC tumors, +/- anti-PD-1 antibodies (scale bar = 10 mm). s. Liver-to-body weight ratios from panel r . Each data point is one liver and P values were determined by one-way ANOVA followed by Tukey’s multiple comparisons test. For all data in this figure, error bars show mean ± s.e.m and statistical tests are stated above. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001; ns, not significant.

Article Snippet: The membranes were blocked with 5%(w/v) skim milk (BD Bioscience, USA) for 1 h at room temperature, and then incubated with the following primary antibodies overnight at 4 °C: TYK2 (Proteintech #16412-1-AP, 1:1000), TYK2 (Cell Signaling Technology #35615S, 1:1000), Phospho-TYK2 (Cell Signaling Technology, 68790S, 1:1000), STAT1 (Cell Signaling Technology, 14994S, 1:1000), Phospho-STAT1 (Cell Signaling Technology, 9167S, 1:1000), STAT3 (Cell Signaling Technology, 9139T, 1:1000), Phospho-STAT3 (Cell Signaling Technology, 9145S, 1:1000) and β-actin (Cell Signaling #4970, 1:1000).

Techniques: Flow Cytometry, Two Tailed Test

Journal: bioRxiv

Article Title: Somatic TYK2 activating mutations in tumor-infiltrating T cells promote anti-cancer immunity

doi: 10.1101/2025.10.13.682217

Figure Lengend Snippet: Tyk2 D807V mutant mice showed increased T cell infiltration upon tumor challenge. a. Representative flow cytometry gating strategy for immune cell subset identification from tumor-infiltrating CD45⁺ cells.

Article Snippet: The membranes were blocked with 5%(w/v) skim milk (BD Bioscience, USA) for 1 h at room temperature, and then incubated with the following primary antibodies overnight at 4 °C: TYK2 (Proteintech #16412-1-AP, 1:1000), TYK2 (Cell Signaling Technology #35615S, 1:1000), Phospho-TYK2 (Cell Signaling Technology, 68790S, 1:1000), STAT1 (Cell Signaling Technology, 14994S, 1:1000), Phospho-STAT1 (Cell Signaling Technology, 9167S, 1:1000), STAT3 (Cell Signaling Technology, 9139T, 1:1000), Phospho-STAT3 (Cell Signaling Technology, 9145S, 1:1000) and β-actin (Cell Signaling #4970, 1:1000).

Techniques: Mutagenesis, Flow Cytometry

Journal: bioRxiv

Article Title: Somatic TYK2 activating mutations in tumor-infiltrating T cells promote anti-cancer immunity

doi: 10.1101/2025.10.13.682217

Figure Lengend Snippet: Tyk2 D807V reduces tumor burden in vivo through CD8⁺ T cells. a. Syngeneic MC38 growth in Tyk2 D807V het or WT littermates, with anti-NK1.1 or anti-IgG antibody treatments. b. Tumor weights from panel a . c. Syngeneic MC38 growth in Tyk2 D807V het or WT littermates, with anti-F4/80 or anti-IgG antibody treatments. d. Tumor weights from panel c . e. Syngeneic MC38 growth in Tyk2 D807V het or WT littermates, with anti-Ly6G or anti-IgG antibody treatments. f. Tumor weights from panel e . g. Syngeneic MC38 growth in Tyk2 D807V het or WT littermates, with anti-CD3 or anti-IgG antibody treatments. h. Tumor weights from panel g . i. Syngeneic MC38 growth in Tyk2 D807V het or WT littermates, with anti-CD4 or anti-IgG antibody treatments. j. Tumor weights from panel i . k. Syngeneic MC38 growth in Tyk2 D807V het or WT littermates, with anti-CD8 or anti-IgG antibody treatments. l. Tumor weights from panel k . m. Syngeneic MC38 growth in mice transplanted with CD8⁺ T cells from Tyk2 D807V het or WT littermates. n. Tumor weights from panel m . o. Representative tumors from panel m . p. Syngeneic RIL175 growth in mice transplanted with CD8⁺ T cells from Tyk2 D807V het or WT littermates. q. Tumor weights from panel p . r. Representative tumors from panel p . s. Syngeneic B16 growth in mice transplanted with CD8⁺ T cells from Tyk2 D807V het or WT littermates. t. Tumor weight collected from panel s . u. Representative tumors from panel s . All data in this figure are presented as mean ± s.e.m. Tumor cells were subcutaneously injected into both flanks of each mouse, so each mouse bears two tumors. Each data point represents an individual tumor (N). For syngeneic models growth curves (panels a,c,e,g,i,k,m,p,s ), P values were determined by two-way ANOVA. For tumor weight panels, P values were determined by one-way ANOVA followed by Tukey’s multiple comparisons test (panels b,d,f,h,j,i ), or two-tailed unpaired t -tests (panels n,q,t ). * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001; ns, not significant. Of note, the data in panels a - h share the same WT+anti-IgG and het+anti-IgG control groups. And the data in panels i-l share the same WT+anti-IgG and het+anti-IgG control groups. For all gross images, tumors from each group were ranked by size in descending order, and the top six are shown.

Article Snippet: The membranes were blocked with 5%(w/v) skim milk (BD Bioscience, USA) for 1 h at room temperature, and then incubated with the following primary antibodies overnight at 4 °C: TYK2 (Proteintech #16412-1-AP, 1:1000), TYK2 (Cell Signaling Technology #35615S, 1:1000), Phospho-TYK2 (Cell Signaling Technology, 68790S, 1:1000), STAT1 (Cell Signaling Technology, 14994S, 1:1000), Phospho-STAT1 (Cell Signaling Technology, 9167S, 1:1000), STAT3 (Cell Signaling Technology, 9139T, 1:1000), Phospho-STAT3 (Cell Signaling Technology, 9145S, 1:1000) and β-actin (Cell Signaling #4970, 1:1000).

Techniques: In Vivo, Injection, Two Tailed Test, Control

Journal: bioRxiv

Article Title: Somatic TYK2 activating mutations in tumor-infiltrating T cells promote anti-cancer immunity

doi: 10.1101/2025.10.13.682217

Figure Lengend Snippet: The anti-tumor effects of TYK2 D807V mice are mediated by CD8⁺ T cells. a. MC38 syngeneic tumors from het Tyk2 D807V and WT littermates treated with anti-IgG, anti-CD3ε, anti-NK1.1, anti-F4/80 and anti-Ly6G antibodies. Tumors from each group were ranked by size in descending order, and the top six are shown. b. MC38 syngeneic tumors from het Tyk2 D807V and WT littermates treated with anti-IgG, anti-CD4 and anti-CD8β antibodies. Tumors from each group were ranked by size in descending order, and the top six are shown. c. Experimental schema for adoptive transplantation of CD8⁺ T cells.

Article Snippet: The membranes were blocked with 5%(w/v) skim milk (BD Bioscience, USA) for 1 h at room temperature, and then incubated with the following primary antibodies overnight at 4 °C: TYK2 (Proteintech #16412-1-AP, 1:1000), TYK2 (Cell Signaling Technology #35615S, 1:1000), Phospho-TYK2 (Cell Signaling Technology, 68790S, 1:1000), STAT1 (Cell Signaling Technology, 14994S, 1:1000), Phospho-STAT1 (Cell Signaling Technology, 9167S, 1:1000), STAT3 (Cell Signaling Technology, 9139T, 1:1000), Phospho-STAT3 (Cell Signaling Technology, 9145S, 1:1000) and β-actin (Cell Signaling #4970, 1:1000).

Techniques: Transplantation Assay

Journal: bioRxiv

Article Title: Somatic TYK2 activating mutations in tumor-infiltrating T cells promote anti-cancer immunity

doi: 10.1101/2025.10.13.682217

Figure Lengend Snippet: Tyk2 D807V also impairs cancer growth in a cell-autonomous manner. a. Cell viability assay (CCK-8) performed on NRAS G12V / shp53 HCC cells isolated from WT and Tyk2 D807V het mice grown in culture ( n = 3,3). P values were determined by two-way ANOVA. a. b. NRAS G12V / shp53 HCC cells isolated from WT and Tyk2 D807V het mice were implanted into WT C57/B6 mice ( n = 26 Tyk2 WT and 22 Tyk2 D807V het tumors). b. Tumor weights from panel b . P values were determined using two-tailed unpaired t -test. c. Representative tumors from panel b . Tumors were ranked by size in descending order, and the top six are shown. d. Western blot of H2.35 cells overexpressing sg TYK2 knockout vectors. e. Transposon constructs containing sgRNA- NRAS G12V , shp53 , and a plasmid containing SB100 transposase. f. Representative livers of sg Tyk2 and sgCtrl with shp53/ NRAS G12V tumors (scale bar = 10 mm). g. Liver-to-body weight ratios from panel g . Each data point is one liver and P values were determined by two-tailed unpaired t -test. h. Surface tumor numbers from panel g . Each data point is one liver and P values were determined by two-tailed unpaired t -test. All data in this figure are presented as mean ± s.e.m. Tumor cells were subcutaneously injected into both flanks of each mouse, so each mouse bears two tumors. Each data point represents an individual tumor (N). * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001; ns, not significant.

Article Snippet: The membranes were blocked with 5%(w/v) skim milk (BD Bioscience, USA) for 1 h at room temperature, and then incubated with the following primary antibodies overnight at 4 °C: TYK2 (Proteintech #16412-1-AP, 1:1000), TYK2 (Cell Signaling Technology #35615S, 1:1000), Phospho-TYK2 (Cell Signaling Technology, 68790S, 1:1000), STAT1 (Cell Signaling Technology, 14994S, 1:1000), Phospho-STAT1 (Cell Signaling Technology, 9167S, 1:1000), STAT3 (Cell Signaling Technology, 9139T, 1:1000), Phospho-STAT3 (Cell Signaling Technology, 9145S, 1:1000) and β-actin (Cell Signaling #4970, 1:1000).

Techniques: Viability Assay, CCK-8 Assay, Isolation, Two Tailed Test, Western Blot, Knock-Out, Construct, Plasmid Preparation, Injection

Journal: bioRxiv

Article Title: Somatic TYK2 activating mutations in tumor-infiltrating T cells promote anti-cancer immunity

doi: 10.1101/2025.10.13.682217

Figure Lengend Snippet: Pharmacological inhibition of TYK2 promotes tumor growth. a. Representative images of A549 tumor co-cultured with primary T cells after treatment with Deucravacitinib or vehicle, scale bars = 1000 μm ( n= 3 replicates per group). b. Relative tumor area of A549 tumor co-cultured with primary T cells after treatment with Deucravacitinib or vehicle ( n =3 replicates per group). c. IFNγ levels measured in A549 tumor cells co-cultured with primary T cells after treatment with Deucravacitinib or vehicle ( n =3 replicates per group). d. Syngeneic MC38 growth in mice receiving deucravacitinib ( n =16 tumors) or vehicle control ( n =14 tumors). e. Tumor weight collected from panel d . f. Representative IHC images of tumor infiltrating CD8⁺ cells in MC38 implanted tumors receiving deucravacitinib or vehicle. (scale bars = 150 μm) g. Quantification of tumor infiltrating CD8⁺ cell area of panel f ( n =10,10 for vehicle, deucravacitinib treated tumors). h. Syngeneic RIL175 tumor growth in mice receiving deucravacitinib ( n =14 tumors) or vehicle control ( n =16 tumors). i. Tumor weight collected from panel h . j. Representative syngeneic tumors from panel h . Tumors from each group were ranked by size in descending order, and the top six were selected for imaging. k. Syngeneic B16 tumor growth in mice receiving deucravacitinib ( n =12 tumors) or vehicle control ( n =12 tumors). l. Tumor weight collected from panel k . m. Representative syngeneic tumors from panel k . Tumors from each group were ranked by size in descending order, and the top six were selected for imaging. n. Syngeneic MC38 growth in Tyk2 D807V het or WT littermates, with vehicle or deucravacitinib treatments. o. Tumor weights from panel n . p. Representative syngeneic tumors from panel n . Tumors from each group were ranked by size in descending order, and the top six were selected for imaging. All data in this figure are presented as mean ± s.e.m. Tumor cells were subcutaneously injected into both flanks of each mouse, so each mouse bears two tumors. Each data point represents an individual tumor (N). For syngeneic models growth curves (panels d,h,k,n ), P values were determined by two-way ANOVA. For panels e,g,i,l,o , P values were determined by two-tailed unpaired t -tests. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001; ns, not significant.

Article Snippet: The membranes were blocked with 5%(w/v) skim milk (BD Bioscience, USA) for 1 h at room temperature, and then incubated with the following primary antibodies overnight at 4 °C: TYK2 (Proteintech #16412-1-AP, 1:1000), TYK2 (Cell Signaling Technology #35615S, 1:1000), Phospho-TYK2 (Cell Signaling Technology, 68790S, 1:1000), STAT1 (Cell Signaling Technology, 14994S, 1:1000), Phospho-STAT1 (Cell Signaling Technology, 9167S, 1:1000), STAT3 (Cell Signaling Technology, 9139T, 1:1000), Phospho-STAT3 (Cell Signaling Technology, 9145S, 1:1000) and β-actin (Cell Signaling #4970, 1:1000).

Techniques: Inhibition, Cell Culture, Control, Imaging, Injection, Two Tailed Test