mouse cytokine array g3 kit  (RayBiotech inc)

Journal: Molecular Cancer

Article Title: Novel induction of CD40 expression by tumor cells with RAS/RAF/PI3K pathway inhibition augments response to checkpoint blockade

doi: 10.1186/s12943-021-01366-y

Figure Lengend Snippet: Rigosertib induces CD40 on melanoma cells and promotes the anti-tumor immunity. a , b Cytokine array of YUMM3.3 tumor lysate samples at day 17 post-treatment. c Live cells were concatenated after downsampling to ~ 20,000 events for t-SNE analysis through flow cytometry. d Flow cytometric analysis of YUMM3.3 tumors in ROSA reporter mice at day 14 post RGS treatment. e Mean fluorescence intensity (MFI) on the CD45 − CD40 + cells isolated from day 17 YUMM3.3 tumors in C57BL/6 mice. f , g Melanoma cells were treated with indicated drugs for 48 h and CD40 expression was detected by flow cytometry. h , i Viability of YUMM3.3 and B16F10 cells was examined by flow cytometry. j Flow cytometric analysis of CD40 expression in response to treatment with IFNγ (500 U/ml, 48 h) in different clones of YUMM3.3 cells where shRNAs targeted coding sequence (CDS), 3″ untranslated region (UTR), or random sequences (Ctrl). k Tumor volume of CD40 knockdown clones in C57BL/6 mice was determined ( n ≥ 4 mice per group). l Tumor weight and tumor volume of YUMM3.3 cells growing in C57BL/6-CD40 knockout male and female mice treated with RGS (300 mg/kg). a - e data were replicates from one experiment ( n = 5 ~ 10 per group). f - l pooled data obtained from at least two different experiments ( n = 3 ~ 6) are shown

Article Snippet: Tumor lysates, serum, and cell culture supernatant samples were examined in cytokine arrays using Mouse Cytokine Array G3 Kit (62 cytokines) (RayBiotech), per the manufacture’s protocol.

Techniques: Flow Cytometry, Fluorescence, Isolation, Expressing, Clone Assay, Sequencing, Knock-Out