Chapter 4 104 Alexa488: 0.9987; R2-PE: 0.9948; R2-APC: 0.9955) indicated that obtained MFI could be readily converted into MESF values. EV-Track Experimental details have been uploaded to the EV-TRACK (ID: EV220008), an open-access knowledgebase recommended by the International Society for Extracellular Vesicles to track worldwide EV research.1,30 Statistical Analysis GraphPad Prism 8.0 (GraphPad Software, USA) was used to analyze and visualize the data. Data were presented as mean ± standard deviation. In the calibration of MESF in IFCM, the analysis templates calculated the correlation coefficient (R2) values between MFI and MESF. The coefficient of variation (CV) reflected the reproducibility of IFCM. Paired t-tests were used to show the differences in uEVs numbers or urinary pH between urine samples with different treatments. Unpaired t-tests were utilized to show the differences in urinary protein levels between HCs and KTRs. Linear regression analysis demonstrated the association between IFCM and TR-FIA readouts. Statistical significance was defined by p-values < 0.05 (twotailed). ACKNOWLEDGMENTS We thank the support from the China Scholarship Council (No. 202008430154) and Wenda Verschoor and Rens Kraaijeveld for helping solve the technical questions of IFCM and structure this paper. We acknowledge Natasja Dits at the Department of Urology (Erasmus Medical Center; EMC) for the TEM images and Manou van Alphen for the support of TR-FIA. All figures were created with BioRender.com. DISCLOSURE G. Jenster has a license agreement with Cell GS for the CD9 and CD63 TR-FIA (TRIFic). Other author reports no conflicts of interest in this work.
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