fpr3 taqman genexpression assay  (Thermo Fisher)

Journal: Cell Reports Medicine

Article Title: Myeloid cells coordinately induce glioma cell-intrinsic and cell-extrinsic pathways for chemoresistance via GP130 signaling

doi: 10.1016/j.xcrm.2024.101658

Figure Lengend Snippet: GP130 is essential for humanin-induced chemoresistance (A) Quantitative reverse-transcription PCR (RT-PCR) for the humanin receptor subunit IL6ST (encoding GP130) was performed; note that IL6ST levels are much higher in humanin-sensitive than humanin-insensitive hGBMs. (B) hGBMs were stimulated with HN or HNG, partly sc144 was coapplied, which consistently abrogated the protumorigenic effect of HN and HNG (dashed line: controls without HN or sc144). (C) Expansion of hGBM1-HN-WT, HN-C8A, or HN-L9R cells, with or without sc144. (D) Humanin expression levels in brain slices with hiPSC-derived microglia and hGBM1 cells were attenuated after addition of sc144 (graphically summarized in E). The number of biological replicates is indicated (dots in graphs indicate data from individual experiments); all error bars are presented as mean ± SDM. Statistical significance is shown by one-way ANOVA in (A) and t test in (B–D): ∗∗ p < 0.01; ∗∗∗∗ p < 0.0001; NS, not significant.

Article Snippet: IL6ST TaqMan Genexpression Assay , ThermoFisher Scientific , Cat#: Hs00174360_m1.

Techniques: Reverse Transcription, Reverse Transcription Polymerase Chain Reaction, Expressing, Derivative Assay

Journal: Cell Reports Medicine

Article Title: Myeloid cells coordinately induce glioma cell-intrinsic and cell-extrinsic pathways for chemoresistance via GP130 signaling

doi: 10.1016/j.xcrm.2024.101658

Figure Lengend Snippet: Humanin-mediated BTB formation is blunted by GP130 blockage increasing chemotherapeutic efficacy and survival (A) Quantification of vascular mural coverage on tumor vessels in orthotopic HN-WT or HN-C8a GBMs with or without TMZ (indicated by blue bars). (B and C) Vascular mural coverage of tumor vessels in TMZ-treated orthotopic HN-WT GBMs with or without BZA. (D) Endothelial cells and pericytes were purified from a transgenic GBM mouse model (GL261, n = 11; infused with humanin, HN, or aCSF, control) and analyzed by transcriptomics; GSEA indicated enrichment for angiogenic traits in HN-stimulated endothelia (as compared to aCSF). (E) Signaling pathways between endothelia and pericytes were analyzed from HN-infused versus control GBMs; in HN-infused GBMs, pericytes promote IL6ST (GP130) signaling in endothelia; in HN-infused GBMs, endothelia promote BMP signaling in pericytes. (F) In HN-WT GBMs, receiving TMZ cotreatment with BZA reduced pericyte (PDGFRB+) coverage of tumor vessels (CD31 + ; as compared to cotreatment with vehicle). (G) Intravenous application of 70 kDA dextran as a tracer for vessel tightness showed that BZA-treated gliomas had significantly increased leakage (across CD31 + vessels) into the tumor parenchyma, as compared to vehicle-treated mice. (H) In mice with orthotopic HN-WT GBMs, intracerebral infusion of sc144 (10 μM) during TMZ chemotherapy prolonged survival as compared to intracerebral infusion of vehicle ( n = 12 per group). (I) Schematic summary: the BTB and DDR protect humanin-sensitive GBMs from TMZ. GP130 inhibitors reduce BTB tightness and blunt chemoresistance. Scale bars indicate 200 μm in (A) and 20 μm in (G). The number of biological replicates is indicated (dots in graphs indicate data from individual mice); all error bars are presented as mean ± SDM. Statistical significance is shown by one-way ANOVA (A), t test (F, G), or by Mantel-Cox test (F): ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: IL6ST TaqMan Genexpression Assay , ThermoFisher Scientific , Cat#: Hs00174360_m1.

Techniques: Purification, Transgenic Assay, Control, Protein-Protein interactions

Journal: Cell Reports Medicine

Article Title: Myeloid cells coordinately induce glioma cell-intrinsic and cell-extrinsic pathways for chemoresistance via GP130 signaling

doi: 10.1016/j.xcrm.2024.101658

Figure Lengend Snippet:

Article Snippet: IL6ST TaqMan Genexpression Assay , ThermoFisher Scientific , Cat#: Hs00174360_m1.

Techniques: Plasmid Preparation, Recombinant, Transfection, Fluorescence, Staining, Reverse Transcription, Gene Expression, Liposomes, Mutagenesis, shRNA, Control, Construct, Software, Imaging, Functional Assay, Dissection, Sequencing, Real-time Polymerase Chain Reaction