single cell suspension tumor isolation  (Miltenyi Biotec)

Journal: bioRxiv

Article Title: STING activation promotes robust immune response and tumor regression in glioblastoma models

doi: 10.1101/2022.02.28.481908

Figure Lengend Snippet: a , Immunoblot of cGAS, STING, TBK1, IRF-3 and GAPDH levels in patient GBM tissue, patient-derived GBM cells and murine GL261 and CT-2A glioma cells. b , Quantification of tissue microarray (TMA) sections for STING and pTBK1, as percentage of positive pixels. AA, n = 77; GBM, n = 49; oligo, n = 10; brain, n = 16. P values calculated by oneway ANOVA. c , Representative TMA sections immunostained for STING and pTBK1 in normal brain and d , GBM. Scale bars = 400 and 100 μm. e , Immunofluorescence staining showing partial activation of the STING cascade in GL261 tumors and the colocalization of pTBK1 within the vasculature (CD31 and α-SMA). Left panel: white arrows point to blood vessels. Right panel: Split channels show colocalization of pTBK1 with the vascular markers CD31 and α-SMA. Scale bar = 100 μm. f , Multiplex immunofluorescence staining on a whole brain section from a GL261 tumor bearing mouse showing DAPI (blue), CD31 (green), STING (yellow) and F4/80 (red). Scale bar = 1 mm. g , h and i , Immunofluorescence staining of the tumor zone showing selected markers as indicated. Scale bars = 400 and 100 μm.

Article Snippet: Single mouse tumor cell suspensions were obtained using a mouse Tumor Disassociation Kit from Miltenyi Biotec (Cat# 130-096-730).

Techniques: Western Blot, Derivative Assay, Microarray, Immunofluorescence, Staining, Activation Assay, Multiplex Assay

Journal: bioRxiv

Article Title: STING activation promotes robust immune response and tumor regression in glioblastoma models

doi: 10.1101/2022.02.28.481908

Figure Lengend Snippet: a, Timeline of the GL261 in vivo experiments for BILs. Biologically independent animals per group (ADU-S100/PBS day 3, n = 3; PBS day 7, n = 3; ADU-S100 day 7, n = 5). P values calculated by two-way ANOVA. b , BIL profile of GL261 tumors at days 3 and 7 using a typical gating procedure. c , Percentage of PD-L1 + CD45 - cells. P value calculated by unpaired t-test. d , G-MDSC populations within the BILs. P value calculated by unpaired t-test. e , 2D t-SNE plots at day 3 and 7, treated mice in red and controls in dark grey. f , t-SNE map for treated mice at day 3 colored by the FlowSOM populations. g, Heatmap and hierarchical clustering of the FlowSOM populations at day 3. h , Immunofluorescence staining on a whole brain section and the necrotic tumor zone 72 h after ADU-S100 treatment (50 μg, bolus in PBS).

Article Snippet: Single mouse tumor cell suspensions were obtained using a mouse Tumor Disassociation Kit from Miltenyi Biotec (Cat# 130-096-730).

Techniques: In Vivo, Immunofluorescence, Staining

Journal: bioRxiv

Article Title: STING activation promotes robust immune response and tumor regression in glioblastoma models

doi: 10.1101/2022.02.28.481908

Figure Lengend Snippet: a , BILs profile of CT-2A tumors at days 3 and 7 using a typical gating procedure. Biologically independent animals per group (PBS, n = 3; ADU-S100, n = 4). P values calculated by two-way ANOVA. b , PD-L1 + percentage of CD45 - cells. P value calculated by unpaired t-test. c , MDSC populations within the BILs. P values calculated by one-way ANOVA. d , 2D t-SNE plots at day 3, treated mice in red and controls in dark grey. e , t-SNE map for treated mice at day 3 colored by the FlowSOM populations; main upregulated FlowSOM populations are highlighted. f, Heatmap and hierarchical clustering of the FlowSOM populations at day 3.

Article Snippet: Single mouse tumor cell suspensions were obtained using a mouse Tumor Disassociation Kit from Miltenyi Biotec (Cat# 130-096-730).

Techniques: