Journal: Matrix biology : journal of the International Society for Matrix Biology
Article Title: Targeting the αVβ3/NgR2 pathway in neuroendocrine prostate cancer
doi: 10.1016/j.matbio.2023.11.003
Figure Lengend Snippet: M1 and M2 marker expression is not affected by incubation with sEVs derived from healthy donors or PrCa patients. Flow cytometric analysis of monocytes isolated from healthy volunteers. ( A ) Histogram representation of the mean fluorescent intensity (MFI) of M2 polarization markers (CD163 and CD204, top panels) and M1 polarization markers (CD80 and HLA-DR, bottom panels) expressed by monocytes after incubation with PBS (−, green) or PrCa patient plasma-derived sEVs (orange) ( n = 13). The isotype control is depicted in red. ( B ) Statistical analysis of M2 marker (CD163 and CD204, top panels) and M1 marker (CD80 and HLA-DR, bottom panels) expression in monocytes after incubation with PBS (−) or PrCa patient plasma-derived sEVs ( n = 13). ( C ) Histogram representation of the MFI of M2 (top panels) and M1 (bottom panels) markers expressed by monocytes after incubation with PBS (−, green), healthy donor (Healthy d.) plasma-derived sEVs (blue), or PrCa patient plasma-derived sEVs (orange) ( n = 3). The isotype control is depicted in red. ( D ) Statistical analysis of M2 (top panels) and M1 (bottom panels) marker levels in monocytes after incubation with PBS (−), healthy donor plasma-derived sEVs or PrCa patient plasma-derived sEVs ( n = 3). ( B, D ) The results are shown in scatter graphs. Paired t -test was used for paired two group comparison; ANOVA for repeated measures with Tukey’s multiple comparisons test was used for multiple groups comparisons with matched samples. SAS 9.4 and Prism 7 were used for data analysis. A.U.: Arbitrary Units.
Article Snippet: Flow cytometry experiments were conducted using the following mouse Abs to human: Alexa Fluor 488 conjugated CD14 (BD Biosciences, 562,689); for M2 polarization phenotyping: APC-conjugated CD163 (Miltenyi, 130–112–129 or Biolegend, 333,610), PE conjugated CD204 (BD Biosciences, 566,251); for M1 polarization phenotyping: APC conjugated CD80 (Biolegend, 305,220), PE-conjugated HLA-DR (BD Biosciences, 555,812); for isotype control: Alexa Fluor 488 conjugated IgG2b,κ isotype control (BD Biosciences, 558,716), APC conjugated REA control antibody IgG1, REAfinity TM (Miltenyi, 130–113–446) or APC conjugated IgG1,κ isotype control antibody (Biolegend, 400,120) and PE conjugated IgG1,κ isotype control (BD Biosciences, 551,436).
Techniques: Marker, Expressing, Incubation, Derivative Assay, Isolation, Clinical Proteomics, Control, Comparison
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