Were processed and imaged identically. Inserts were imaged using confocal microscopy. Scale bar (150 mm). Inset scale bar (50 mm). (DOCX) Figure S4 Histogram plots from antigens in Table 2. Antigens with increase in percent BIRB 796 cost positivity by at least 2-fold. Plot in red is corresponding isotype control. Blue line represents reactivity for the specified antibody. (DOCX) Figure S5 Histogram plots from antigens in Table 3. Antigens with decrease in percent positivity by at least 2-fold. Plot in red is corresponding isotype control. Blue line represents reactivity for the specified antibody. (DOCX) Figure S6 FACS plots from stem cell marker analysis inTable 4. Left: Histogram plots of EpCAM staining for the indicated cell lines. Red line indicates isotype control. Black line is reactivity for EpCAM antibody. Right: EpCAM+ cells from histogram gates shown on left stained with CD133-APC (y-axis) and CD44-PE (x-axis). (DOCX)Figure S7 CD44 antigen sensitivity to enzymatic detachment. Enzymatic treatment affects antigen expression. The HCT116 cell line was enzymatically detached from the tissue culture plate using either trypsin (TryPLE, left) or papain (right) prior to standard FACS Danusertib antibody labeling and analysis. The expression of CD44 was virtually eliminated after papain treatment, suggesting enzymatic cleavage of this epitope. (DOCX) Table S1 Complete SW480 profiling results.Supporting InformationFigure S1 Complete antibody array results. Complete results from barcoded antibody array screening of SW480, SW620, and HCT116 CRC cell lines. The position of each antibody on the plate is indicated from rows A and columns 1?12. To generate a heatmap of the expression of antigens,(XLSX)Multiplexed FACS Antibody Array in Colon CancerTable S2 Complete SW620 profiling results.(XLSX)Table S3 Complete HCT116 profiling results.of percent cell positivity in Tables 2 and 3. Discordance is indicated with an asterisk in Tables 2 and 3. (XLSX)(XLSX) Calculation and comparison of mean fluorescence intensities of SW480 and SW620 cells. Median, mean, and normalized fluorescence intensities derived from Tables S1 and S2. Fold change differences in SW480 and SW620 are calculated. Green shading: antigen was two-fold increased (SW620/ SW480.2) by mean fluorescence intensity. Red shading: antigen was two-fold decreased (SW620/SW480,0.5). This list was then cross-referenced with the list of antigens identified by comparisonTable SAcknowledgmentsWe appreciate critical insight and manuscript review by members of the Rich laboratory and Dr. Jennifer Yu.Author ContributionsConceived and designed the experiments: KS RIP JGV JDL. Performed the experiments: KS RIP JGV AJ JDL MR. Analyzed the data: KS RIP JGV ABH JDL JNR DRC. Contributed reagents/materials/analysis tools: RIP JGV JE JM MFK CTC RB DRC. Wrote the paper: KS JDL JNR.
Essential tremor (ET) is among the most prevalent movement disorders [1]. In postmortem studies, degenerative changes in the cerebellum, including an increase in the number of Purkinje cell (PC) axonal torpedoes and PC loss have been reported [2,3]. Other pathological features have also been reported in ET, including an increase in the numbers of heterotopic PCs, an increased density of the basket cell axonal plexus surrounding PCs, and Bergmann gliosis [4?]. In contrast, granule cells and parallel fibers seem to be relatively preserved in ET [7]. Whether ET is a neurodegenerative disease is under active discussion [8]. Since PC loss has been reported i.Were processed and imaged identically. Inserts were imaged using confocal microscopy. Scale bar (150 mm). Inset scale bar (50 mm). (DOCX) Figure S4 Histogram plots from antigens in Table 2. Antigens with increase in percent positivity by at least 2-fold. Plot in red is corresponding isotype control. Blue line represents reactivity for the specified antibody. (DOCX) Figure S5 Histogram plots from antigens in Table 3. Antigens with decrease in percent positivity by at least 2-fold. Plot in red is corresponding isotype control. Blue line represents reactivity for the specified antibody. (DOCX) Figure S6 FACS plots from stem cell marker analysis inTable 4. Left: Histogram plots of EpCAM staining for the indicated cell lines. Red line indicates isotype control. Black line is reactivity for EpCAM antibody. Right: EpCAM+ cells from histogram gates shown on left stained with CD133-APC (y-axis) and CD44-PE (x-axis). (DOCX)Figure S7 CD44 antigen sensitivity to enzymatic detachment. Enzymatic treatment affects antigen expression. The HCT116 cell line was enzymatically detached from the tissue culture plate using either trypsin (TryPLE, left) or papain (right) prior to standard FACS antibody labeling and analysis. The expression of CD44 was virtually eliminated after papain treatment, suggesting enzymatic cleavage of this epitope. (DOCX) Table S1 Complete SW480 profiling results.Supporting InformationFigure S1 Complete antibody array results. Complete results from barcoded antibody array screening of SW480, SW620, and HCT116 CRC cell lines. The position of each antibody on the plate is indicated from rows A and columns 1?12. To generate a heatmap of the expression of antigens,(XLSX)Multiplexed FACS Antibody Array in Colon CancerTable S2 Complete SW620 profiling results.(XLSX)Table S3 Complete HCT116 profiling results.of percent cell positivity in Tables 2 and 3. Discordance is indicated with an asterisk in Tables 2 and 3. (XLSX)(XLSX) Calculation and comparison of mean fluorescence intensities of SW480 and SW620 cells. Median, mean, and normalized fluorescence intensities derived from Tables S1 and S2. Fold change differences in SW480 and SW620 are calculated. Green shading: antigen was two-fold increased (SW620/ SW480.2) by mean fluorescence intensity. Red shading: antigen was two-fold decreased (SW620/SW480,0.5). This list was then cross-referenced with the list of antigens identified by comparisonTable SAcknowledgmentsWe appreciate critical insight and manuscript review by members of the Rich laboratory and Dr. Jennifer Yu.Author ContributionsConceived and designed the experiments: KS RIP JGV JDL. Performed the experiments: KS RIP JGV AJ JDL MR. Analyzed the data: KS RIP JGV ABH JDL JNR DRC. Contributed reagents/materials/analysis tools: RIP JGV JE JM MFK CTC RB DRC. Wrote the paper: KS JDL JNR.
Essential tremor (ET) is among the most prevalent movement disorders [1]. In postmortem studies, degenerative changes in the cerebellum, including an increase in the number of Purkinje cell (PC) axonal torpedoes and PC loss have been reported [2,3]. Other pathological features have also been reported in ET, including an increase in the numbers of heterotopic PCs, an increased density of the basket cell axonal plexus surrounding PCs, and Bergmann gliosis [4?]. In contrast, granule cells and parallel fibers seem to be relatively preserved in ET [7]. Whether ET is a neurodegenerative disease is under active discussion [8]. Since PC loss has been reported i.
