proteome profiler human cytokine array kit 9  (R&D Systems)

Journal: Cancer Research

Article Title: Transfer of Damaged Mitochondria from Cancer Cells to Cancer-Associated Fibroblasts Promotes Tyrosine Kinase Inhibitor Tolerance in EGFR -Mutant Lung Cancer

doi: 10.1158/0008-5472.CAN-25-0433

Figure Lengend Snippet: Recruitment of RGS5 + MYL9 + CAFs by DTP cells via CCL11 secretion. A and B, Cytokine arrays and table display changes in cytokine levels from HCC827 cells cultured with various CAF subsets. C, ELISA results reveal concentrations of IL1α, IL13, IL15, MCP-1, MCP-2, CCL11, and GDNF present in the media from HCC827 cells cocultured with different CAF subsets. D, ELISA analysis shows levels of IL13 and CCL11 in media from HCC827 cells cultured with RGS5 + MYL9 + CAFs under vehicle or mitoTEMPO treatment conditions. E, Relative cell viability data for RGS5 + MYL9 + CAFs cocultured with HCC827 cells treated with either vehicle or mitoTEMPO. Osi, osimertinib. F, Schematic overview outlines the migration assay for RGS5 + MYL9 + CAFs. G, Representative images and quantification demonstrate the migration of RGS5 + MYL9 + CAFs toward cocultured HCC827 cells. H, Schematic overview depicts the migration assay for HCC827 cells alongside representative images and quantification of their movement toward cocultured RGS5 + MYL9 + CAFs. I and J, Representative images showing cocultures of RFP-labeled HCC827 cells with GFP-labeled CAFs. Scale bar, 50 μm. K–P, Both HCC827 and RGS5 + MYL9 + CAFs were xenografted into mice treated with osimertinib (5 mg/kg/day) or αCCL11 (2 mg/kg/day). L, Tumor volumes for xenografted mice. M, Body weight of tumors from xenografted mice. N, Percentage survival rate for mice bearing xenografts derived from HCC827 and RGS5 + MYL9 + CAFs. The tick marks in the Kaplan–Meier curves denote censored events corresponding to predefined humane endpoints. These include tumor volume reaching or exceeding 2,000 mm 3 (as measured by caliper) or a loss of body weight greater than 20% from baseline. O, Histologic examinations, including H&E and Ki67 staining, are conducted on tumor tissues from xenograft mice. Scale bar, 100 μm. P, Representative images illustrate TUNEL staining alongside infiltration patterns of RGS5 + MYL9 + CAFs in tumors from xenograft models. Scale bar, 100 μm. The primary CAF subsets utilized in this figure were isolated from patients 07 to 09. Relevant clinical characteristics of these patients are summarized in Supplementary Table S1. Data are presented as the mean ± SD. D and E, Data were analyzed via independent sample t tests; G and M , data were analyzed using one-way ANOVA followed by Dunnett test; N , data were evaluated using a two-sided log-rank test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant.

Article Snippet: Cytokine profiles from LUAD cells were analyzed using the Proteome Profiler Human XL Cytokine Array Kit (RayBio).

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, Migration, Labeling, Derivative Assay, Staining, TUNEL Assay, Isolation