recombinant mouse ccl9  (MedChemExpress)

Journal: Journal for immunotherapy of cancer

Article Title: Tumor-intrinsic TTLL12 drives resistance to cancer immunotherapy via modulating myeloid-derived suppressor cells.

doi: 10.1136/jitc-2024-010873

Figure Lengend Snippet: Figure 6 TTLL12 promotes chemokine CCL9 secretion in colorectal cancer. (A) The workflow to identify the cytokines bridging the connection between tumor TTLL12 and MDSCs in the tumor microenvironment. (B) The heatmap of 33 candidate proteins with p<0.01 and secretory ability predicted by the Human Protein Atlas database. (C) ELISA analysis in the supernatant of CT26 shNC and CT26 shTTLL12 cells. Values are represented as mean±SEM. P values were determined by two-tailed t-tests, ***p<0.001. (D) qRT-PCR analysis of CT26 shNC and CT26 shTTLL12 cells. Values are represented as mean±SEM. P values were determined by two-tailed t-tests, ***p<0.001. (E) Tumor image from the immunocompetent Balb/c mice subcutaneously implanted with CT26-control cells, TTLL12-overexpressing CT26 cells as well as TTLL12-overexpressing CT26 cells with CCL9 silence. (F–G) Tumor volume (F) and tumor weight (G) in Balb/c mice. n=5 mice for each group. Values are represented as mean±SEM. P values calculated by two-way ANOVA, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. (H) Quantification of CCL9 concentration in tumors. Values are represented as mean±SEM. P values calculated by two-way ANOVA, ****p<0.0001. (I) Flow cytometry analysis to test the MDSCs infiltration in tumors. Values are represented as mean±SEM. P values calculated by two-way ANOVA, ***p<0.001, ****p<0.0001. ANOVA, analysis of variance; MDSC, myeloid-derived suppressor cells; qRT-PCR, quantitative reverse transcription-polymerase chain reaction; TTLL12, tubulin tyrosine ligase 12.

Article Snippet: The concentrations of CCL9 in mice were measured by using a Mouse CCL9 ELISA Kit from Boster (Wuhan, China), following the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Two Tailed Test, Quantitative RT-PCR, Control, Concentration Assay, Flow Cytometry, Derivative Assay, Reverse Transcription, Polymerase Chain Reaction

Journal: Journal for immunotherapy of cancer

Article Title: Tumor-intrinsic TTLL12 drives resistance to cancer immunotherapy via modulating myeloid-derived suppressor cells.

doi: 10.1136/jitc-2024-010873

Figure Lengend Snippet: Figure 7 TTLL12 induces CCL9 expression, probably by binding to its promoter region. (A) The intracellular location of TTLL12 was detected by IHC analysis based on colorectal cancer samples in our center. Representative images of IHC staining from three tissue samples were shown. Red arrows: cell nucleus. Scale bars: 50 µm (left panel), 25 µm (right panel). (B–C) Western blot analysis of the cytoplasm and nucleus of CT26 cells transfected with plasmid harboring TTLL12 overexpression or knockdown with TTLL12 antibody, α-tubulin (a cytosolic marker) antibody and Lamin-B (a marker for nuclear compartments) antibody. (D) The promoter region of CCL9 covered by the primer. (E–F) ChIP-PCR assay in DLD1/TTLL12 and CT26/TTLL12 cells. Values are represented as mean±SEM. P values were determined by two-tailed t-tests, ****p<0.0001. ChIP, chromatin iImmunoprecipitation; IHC, immunohistochemistry; TTLL12, tubulin tyrosine ligase 12.

Article Snippet: The concentrations of CCL9 in mice were measured by using a Mouse CCL9 ELISA Kit from Boster (Wuhan, China), following the manufacturer’s instructions.

Techniques: Expressing, Binding Assay, Immunohistochemistry, Western Blot, Transfection, Plasmid Preparation, Over Expression, Knockdown, Marker, Two Tailed Test

Journal: Journal for immunotherapy of cancer

Article Title: Tumor-intrinsic TTLL12 drives resistance to cancer immunotherapy via modulating myeloid-derived suppressor cells.

doi: 10.1136/jitc-2024-010873

Figure Lengend Snippet: Figure 9 Working model of TTLL12-mediated antitumor immune response. Tumor-intrinsic TTLL12 depends on MDSCs to promote tumor progression and knockdown of TTLL12 can enhance the antitumor efficacy of anti-programmed cell death protein 1 therapy in an immunocompetent mouse model. Mechanistically, tumor- derived TTLL12 promotes chemokine CCL9 secretion to modulate MDSCs by inducing the transcriptional expression of CCL9, probably by binding to its promoter region. MDSC, myeloid-derived suppressor cell; NK, natural killer; TTLL12, tubulin tyrosine ligase 12.

Article Snippet: The concentrations of CCL9 in mice were measured by using a Mouse CCL9 ELISA Kit from Boster (Wuhan, China), following the manufacturer’s instructions.

Techniques: Knockdown, Derivative Assay, Expressing, Binding Assay