Journal: Nature communications

Article Title: An Artificial Intelligence-guided signature reveals the shared host immune response in MIS-C and Kawasaki disease.

doi: 10.1038/s41467-022-30357-w

Figure Lengend Snippet: Fig. 5 An integrated analysis of mesoscale (cytokine) data, ViP/sViP transcriptomic signatures and laboratory and clinical parameters reveals features that are unique to MIS-C. a Heatmap displays the results of hierarchical agglomerative clustering of acute KD (KD-AV; n = 10) and MIS-C (n = 10) subjects using the cytokine profiles determined by mesoscale (MSD) and the laboratory features. Source data are provided as a Source Data file. b Violin plots display PLT (platelet) and AEC (absolute eosinophil counts) in KD and MIS-C (unpaired two-sided Student’s t-test used to test significance). c–e Correlation test (two-sided test of the slope of the regression line compared to zero) between AEC and PLT (c; left) and IL15 and PLT (c; right), and MIP1α and PLT (d) and MIP1α and IL15 (e) are shown, and significance was calculated and displayed using GraphPad Prism 9. Significance: ns: non- significant, ****p < 0.0001. See Supplementary Fig. S3 for all possible correlation tests between clinical and cytokine data in KD, MIS-C and COVID-19. f Correlation tests between PLT (left) or AEC (right) on the Y-axis and gene signature scores on the X-axis [either ViP (top), sViP (middle) or a IL15/IL15RA composite (bottom)] were calculated and displayed as scatter plots using python seaborn lmplots with the p-values. The confidence interval around the regression line is indicated with shades. g Schematic summarizing the findings in MIS-C based on laboratory and RNA seq analysis.

Article Snippet: Formalin-fixed, paraffin-embedded heart tissue sections from COVID19 and KD patients were stained anti-human IL15 receptor A polyclonal antibody (11:200 dilution; proteintech®, Rosemont, IL, USA; catalog# 16744- 1-AP) and anti-human IL15 monoclonal antibody (1:10 dilution; Santa Cruz Biotechnology, Inc., Dallas, TX, USA; catalog# sc-8437) after heat-induced antigen retrieval with Tris buffer containing EDTA (pH 9.0).

Techniques: RNA Sequencing

Journal: Nature communications

Article Title: An Artificial Intelligence-guided signature reveals the shared host immune response in MIS-C and Kawasaki disease.

doi: 10.1038/s41467-022-30357-w

Figure Lengend Snippet: Fig. 6 ViP/sViP signatures correlate with two distinct cardiac phenotypes in MIS-C and KD. a Violin plots display the left ventricular ejection functions (LVEF) in KD and MIS-C patients. Statistical significance was determined by unpaired two-sided Student’s t-test. b–d Correlation tests (two-sided test of the slope of the regression line compared to zero) between LVEF (Y-axis) and gene signature scores on the X-axis [either ViP (b), sViP (c), or a IL15/IL15RA composite (d)] are displayed as a scatter plot and significance was calculated and displayed as in Fig. 5f. The confidence interval around the regression line is indicated with shades. e Bar and violin plots show how a IL15/IL15RA compositive score varies between KD samples. The score classifies KD-AV with giant CAAs from control (KD-CV) samples with a ROC AUC 0.95. Welch’s two sample unpaired two-sided t-test is performed on the composite gene signature score to compute the p values. In multi-group setting each group is compared to the first control group and only significant p values are displayed.

Article Snippet: Formalin-fixed, paraffin-embedded heart tissue sections from COVID19 and KD patients were stained anti-human IL15 receptor A polyclonal antibody (11:200 dilution; proteintech®, Rosemont, IL, USA; catalog# 16744- 1-AP) and anti-human IL15 monoclonal antibody (1:10 dilution; Santa Cruz Biotechnology, Inc., Dallas, TX, USA; catalog# sc-8437) after heat-induced antigen retrieval with Tris buffer containing EDTA (pH 9.0).

Techniques: Control

Journal: Nature communications

Article Title: An Artificial Intelligence-guided signature reveals the shared host immune response in MIS-C and Kawasaki disease.

doi: 10.1038/s41467-022-30357-w

Figure Lengend Snippet: Fig. 7 Summary of findings and conclusions. a Summary of datasets used (publicly available prior ones and new original cohorts) to support the conclusions in this work. Numbers in circles denote the number of subjects in each cohort. b Venn diagram displays the major findings from the current work. ViP/sViP signatures, and more specifically, the IL15/IL15RA specific gene induction are shared between patients in all three diagnostic groups. While these signatures are known to be associated with diffuse alveolar damage in the lungs of patients with COVID-1918, it is associated with CAA in KD and with reduction in cardiac muscle contractility in MIS-C. Overlapping features between each entity are displayed.

Article Snippet: Formalin-fixed, paraffin-embedded heart tissue sections from COVID19 and KD patients were stained anti-human IL15 receptor A polyclonal antibody (11:200 dilution; proteintech®, Rosemont, IL, USA; catalog# 16744- 1-AP) and anti-human IL15 monoclonal antibody (1:10 dilution; Santa Cruz Biotechnology, Inc., Dallas, TX, USA; catalog# sc-8437) after heat-induced antigen retrieval with Tris buffer containing EDTA (pH 9.0).

Techniques: Diagnostic Assay

Journal: EBioMedicine

Article Title: AI-guided discovery of the invariant host response to viral pandemics.

doi: 10.1016/j.ebiom.2021.103390

Figure Lengend Snippet: Fig. 6. ViP signatures reveal an interplay between IL15-storm and NK cell dysfunction in fatal COVID-19. (a) Bubble plots of ROC-AUC values (radius of circles are based on the ROC- AUC) demonstrating the direction of gene regulation (Up, red; Down, blue) for the classification based on the 20 gene severe ViP signature (top) and 166 gene ViP signature (bottom) in following datasets. RNASeq data (GSE115203) from PBMCs and sorted NK cells from PBMCs incubated with uninfected A549 cells for 12 hrs compared to infected A549 cells. PBMCs treated with IL15 compared to IL2 (GSE77601). RNASeq analysis of NK cells (GSE89484) treated with GSK-J4 compared to DMSO. Skin tissue in mice (GSE45551) is treated with anti-IL15RB antibody compared to PBS. (b) RNASeq data of NK cells isolated from two donors prior to vaccination compared (left) to days 1, 3, and 7 post-TIV vaccination like panel A. RNASeq data of NK enriched and NK depleted PBMCs from healthy donors compared to 30 day post-vaccination like panel A. (c, d) Heatmap of 20-gene (panel c) and 166- gene (panel d) ViP signatures in tissues collected during rapid autopsies on patients who succumbed to COVID-19. Genes are ranked according to the strength of differential expres- sion (T-test) in lung tissue between normal and infected tissue. (e) Box plots of IL15 and IL15RA in samples from varying severity of COVID-19. (f-h) Violin plots show levels of plasma IL15 in COVID-19 patients stratified by disease acquity (F), by clinical severity (G) and by gender and age (H). Welch’s two sample unpaired t-test is performed to compute the p values. See also Table S5 for patient metadata.

Article Snippet: Tissues were then incubated with rabbit IL15RA polyclonal antibody (1:200 dilution; proteintech , Rosemont, IL, USA; catalog# 16,744 1-AP) for 1.5 h and mouse IL15 monoclonal antibody (1:10 dilution; Santa Cruz Biotechnology, Inc., Dallas, TX, USA; catalog# sc-8437) at room temperature in a humidified chamber and then rinsed with TBS or PBS 3x, 5 min each.

Techniques: Incubation, Infection, Isolation, Clinical Proteomics

Journal: EBioMedicine

Article Title: AI-guided discovery of the invariant host response to viral pandemics.

doi: 10.1016/j.ebiom.2021.103390

Figure Lengend Snippet: Fig. 7. Lung alveolar cells contribute to the IL15 storm in fatal COVID-19. (a) Normal lung tissue obtained during surgical resection (left) or lung tissue obtained during autopsy stud- ies on COVID-19 patients (right) were stained for IL15 and IL15RA. Representative images are shown. Mag = 10X. (b) Violin plots display the intensity of staining for IL15RA (top) and IL15 (bottom), as determined by IHC profiler. (c) Hospital-free days analysis (45 days followup) of COVID-19 patients (GSE157103) limited to males less than 70 years old using the abundance of IL15 transcripts (intermediate and high groups) is displayed as Kaplan-Meier estimates (left) of cumulative probability of discharge and its relationship with days in hospital. (d) Cox-proportional hazard univariate analysis (right; top) of sViP (high vs low) is compared to ViP signature, Interferon Stimulated Gene-signatures (ISG1, PMID:15619625; ISG2, PMID:21478870), age, gender, ICU admission (icu) and mechanical ventilation (mv). Multivariate Cox-proportional hazard analysis (right; bottom) compares the variables that are significant in univariate settings, i.e., sViP, ICU admission (icu) and mechanical ventilation (mv). (e) Top: Schematic displays the workflow for patient blood col- lection and assessment of IL15 levels by mesoscale. Bottom: Bar (top) and violin (bottom) plots for the levels of IL15 cytokine (score = Z score of the log reduced mesoscale concentra- tion data). ROC AUC numbers indicate the strength of classification between patients with critical/fatal disease course vs. those with non-critical infection. (f) Summary of IL15 signaling and the hypothetical role of NK cells in the severity of COVID-19 infections.

Article Snippet: Tissues were then incubated with rabbit IL15RA polyclonal antibody (1:200 dilution; proteintech , Rosemont, IL, USA; catalog# 16,744 1-AP) for 1.5 h and mouse IL15 monoclonal antibody (1:10 dilution; Santa Cruz Biotechnology, Inc., Dallas, TX, USA; catalog# sc-8437) at room temperature in a humidified chamber and then rinsed with TBS or PBS 3x, 5 min each.

Techniques: Staining, Infection

Journal: EBioMedicine

Article Title: AI-guided discovery of the invariant host response to viral pandemics.

doi: 10.1016/j.ebiom.2021.103390

Figure Lengend Snippet: Fig. 8. Validation of ViP signature-guided therapeutic goals. (a-c) The 166-gene ViP signature-was used to classify liver biopsies from HCV-infected patients treated or not with directly acting anti-viral agents. ROC-AUC values are shown below each bar plot unless otherwise stated. (d) 166-gene ViP signature-based classification of blood samples from HIV- infected patients treated with anti-retroviral therapy (ART). (e) The compound EIDD-2801 (MK-4482; 500 mg/kg) or vehicle (Veh) was administered at indicated doses to Golden Syrian hamsters 4 h prior to intranasal infection with SARS-CoV-2. Hamsters were sacrificed on day 5 and lungs we analyzed by RNA sequencing. (f) Bar (top) and violin (bottom) plots using the ViP (left) or sViP (right) signature-based classification of lung samples from hamsters in E and uninfected controls. (g) Schematic showing the experimental design for validating the ViP signatures as useful tools to assess therapeutic efficacy. Uninf, uninfected; Den3 and Anti-CoV-2 indicate SARS-CoV-2 challenged groups that received either a control mAb or the clone CC12.2 of anti-CoV-2 IgG, respectively. (h) Bar (top) and violin (bottom) plots display the 166- and 20-gene ViP signatures in the uninfected and the SARS- CoV-2 challenged groups, treated with control or anti-CoV-2 IgG. (i-k) Lungs harvested from the 3 groups of hamsters were analyzed by H&E and IHC. Representative images are shown in I. Mag = 10X. Bar graphs in J display the abundance of cellularity and infiltrates in the lungs of the 3 groups, as determined by ImageJ. Violin plots in K display the intensity of staining for IL15RA (top) and IL15 (bottom), as determined by IHC profiler.

Article Snippet: Tissues were then incubated with rabbit IL15RA polyclonal antibody (1:200 dilution; proteintech , Rosemont, IL, USA; catalog# 16,744 1-AP) for 1.5 h and mouse IL15 monoclonal antibody (1:10 dilution; Santa Cruz Biotechnology, Inc., Dallas, TX, USA; catalog# sc-8437) at room temperature in a humidified chamber and then rinsed with TBS or PBS 3x, 5 min each.

Techniques: Biomarker Discovery, Infection, Retroviral, RNA Sequencing, Control, Staining

Journal: EBioMedicine

Article Title: AI-guided discovery of the invariant host response to viral pandemics.

doi: 10.1016/j.ebiom.2021.103390

Figure Lengend Snippet: Fig. 6. ViP signatures reveal an interplay between IL15-storm and NK cell dysfunction in fatal COVID-19. (a) Bubble plots of ROC-AUC values (radius of circles are based on the ROC- AUC) demonstrating the direction of gene regulation (Up, red; Down, blue) for the classification based on the 20 gene severe ViP signature (top) and 166 gene ViP signature (bottom) in following datasets. RNASeq data (GSE115203) from PBMCs and sorted NK cells from PBMCs incubated with uninfected A549 cells for 12 hrs compared to infected A549 cells. PBMCs treated with IL15 compared to IL2 (GSE77601). RNASeq analysis of NK cells (GSE89484) treated with GSK-J4 compared to DMSO. Skin tissue in mice (GSE45551) is treated with anti-IL15RB antibody compared to PBS. (b) RNASeq data of NK cells isolated from two donors prior to vaccination compared (left) to days 1, 3, and 7 post-TIV vaccination like panel A. RNASeq data of NK enriched and NK depleted PBMCs from healthy donors compared to 30 day post-vaccination like panel A. (c, d) Heatmap of 20-gene (panel c) and 166- gene (panel d) ViP signatures in tissues collected during rapid autopsies on patients who succumbed to COVID-19. Genes are ranked according to the strength of differential expres- sion (T-test) in lung tissue between normal and infected tissue. (e) Box plots of IL15 and IL15RA in samples from varying severity of COVID-19. (f-h) Violin plots show levels of plasma IL15 in COVID-19 patients stratified by disease acquity (F), by clinical severity (G) and by gender and age (H). Welch’s two sample unpaired t-test is performed to compute the p values. See also Table S5 for patient metadata.

Article Snippet: MATERIALS & REAGENTS ANTIBODIES USED FOR IMMUNOCYTOCHEMISTRY Name Manufacturer Catalog number Dilution factor IL15 (E-4) Santa Cruz sc-8437 1:10 IL15RA Proteintech 16,744 1-AP 1:200 Goat anti-rabbit Vector Laboratories, Burlingame, USA MP-7401 1:500 Goat anti-mouse Vector Laboratories, Burlingame, USA MP-7402 1:500 INSTRUMENTS Leica DMI4000B (Automated Inverted Microscope) Leica Microsystems DMI4000B Power Pressure Cooker XL Tristar Products FisherbrandTM 150 Handheld Homogenizer Fisher Scientific (continued) 15,340,168 SOFTWARE ImageJ https://imagej.nih.gov/ij/index.html GraphPad Prism https://www.graphpad.com/scientific-software/prism/ KITS, ENZYMES, CHEMICALS, AND REAGENTS ELISA MAX Deluxe set Biolegend 435,104 V-PLEX sandwich immunoassays Mesoscale Discovery (MSD) K151A9H-1 Zinc Formalin Fisher Scientific 23 313,096 Xylene VWR XX0060 4 Hematoxylin Sigma-Aldrich Inc MHS1 Ethanol Koptec UN1170 Sodium Citrate Sigma-Aldrich W302600 DAB (10X) Vector Laboratories, Burlingame, USA SK-4105 Hematoxylin Sigma-Aldrich Inc. MO, USA MHS1 Stable Peroxidase substrate buffer (10x) Thermo Fisher 34,062 1:10 3% Hydrogen Peroxide Target 245 07 3628 Horse Serum Vector Labs 30,022 Paraformaldehyde 16% Solution, EM Grade Electron Microscopy Sciences 15,710 100% Methanol Supelco MX0485 Glycine Fisher Scientific BP381 5 Bovine Serum Albumin Sigma-Aldrich A9647 100G Triton-X 100 Sigma-Aldrich X100 500ML Prolong Glass Invitrogen P36984 Nail Polish (Rapid Dry) Electron Microscopy Sciences 72,180 Gill Modified Hematoxylin (Solution II) Millipore Sigma 65,066 85 Goat serum Vector Laboratories MP-7401 Quick-RNA MicroPrep Kit Zymo Research R1051 Quick-RNA MiniPrep Kit Zymo Research R1054 Ethyl alcohol, pure Sigma-Aldrich (continued) E7023 qScript cDNA SuperMix Quanta Biosciences 95,048 OTHER RNase Away Thermo Fisher Scientific 14 375 35 Noyes Spring Scissors - Angled Fine Science Tools 15,013 12

Techniques: Incubation, Infection, Isolation, Clinical Proteomics

Journal: EBioMedicine

Article Title: AI-guided discovery of the invariant host response to viral pandemics.

doi: 10.1016/j.ebiom.2021.103390

Figure Lengend Snippet: Fig. 7. Lung alveolar cells contribute to the IL15 storm in fatal COVID-19. (a) Normal lung tissue obtained during surgical resection (left) or lung tissue obtained during autopsy stud- ies on COVID-19 patients (right) were stained for IL15 and IL15RA. Representative images are shown. Mag = 10X. (b) Violin plots display the intensity of staining for IL15RA (top) and IL15 (bottom), as determined by IHC profiler. (c) Hospital-free days analysis (45 days followup) of COVID-19 patients (GSE157103) limited to males less than 70 years old using the abundance of IL15 transcripts (intermediate and high groups) is displayed as Kaplan-Meier estimates (left) of cumulative probability of discharge and its relationship with days in hospital. (d) Cox-proportional hazard univariate analysis (right; top) of sViP (high vs low) is compared to ViP signature, Interferon Stimulated Gene-signatures (ISG1, PMID:15619625; ISG2, PMID:21478870), age, gender, ICU admission (icu) and mechanical ventilation (mv). Multivariate Cox-proportional hazard analysis (right; bottom) compares the variables that are significant in univariate settings, i.e., sViP, ICU admission (icu) and mechanical ventilation (mv). (e) Top: Schematic displays the workflow for patient blood col- lection and assessment of IL15 levels by mesoscale. Bottom: Bar (top) and violin (bottom) plots for the levels of IL15 cytokine (score = Z score of the log reduced mesoscale concentra- tion data). ROC AUC numbers indicate the strength of classification between patients with critical/fatal disease course vs. those with non-critical infection. (f) Summary of IL15 signaling and the hypothetical role of NK cells in the severity of COVID-19 infections.

Article Snippet: MATERIALS & REAGENTS ANTIBODIES USED FOR IMMUNOCYTOCHEMISTRY Name Manufacturer Catalog number Dilution factor IL15 (E-4) Santa Cruz sc-8437 1:10 IL15RA Proteintech 16,744 1-AP 1:200 Goat anti-rabbit Vector Laboratories, Burlingame, USA MP-7401 1:500 Goat anti-mouse Vector Laboratories, Burlingame, USA MP-7402 1:500 INSTRUMENTS Leica DMI4000B (Automated Inverted Microscope) Leica Microsystems DMI4000B Power Pressure Cooker XL Tristar Products FisherbrandTM 150 Handheld Homogenizer Fisher Scientific (continued) 15,340,168 SOFTWARE ImageJ https://imagej.nih.gov/ij/index.html GraphPad Prism https://www.graphpad.com/scientific-software/prism/ KITS, ENZYMES, CHEMICALS, AND REAGENTS ELISA MAX Deluxe set Biolegend 435,104 V-PLEX sandwich immunoassays Mesoscale Discovery (MSD) K151A9H-1 Zinc Formalin Fisher Scientific 23 313,096 Xylene VWR XX0060 4 Hematoxylin Sigma-Aldrich Inc MHS1 Ethanol Koptec UN1170 Sodium Citrate Sigma-Aldrich W302600 DAB (10X) Vector Laboratories, Burlingame, USA SK-4105 Hematoxylin Sigma-Aldrich Inc. MO, USA MHS1 Stable Peroxidase substrate buffer (10x) Thermo Fisher 34,062 1:10 3% Hydrogen Peroxide Target 245 07 3628 Horse Serum Vector Labs 30,022 Paraformaldehyde 16% Solution, EM Grade Electron Microscopy Sciences 15,710 100% Methanol Supelco MX0485 Glycine Fisher Scientific BP381 5 Bovine Serum Albumin Sigma-Aldrich A9647 100G Triton-X 100 Sigma-Aldrich X100 500ML Prolong Glass Invitrogen P36984 Nail Polish (Rapid Dry) Electron Microscopy Sciences 72,180 Gill Modified Hematoxylin (Solution II) Millipore Sigma 65,066 85 Goat serum Vector Laboratories MP-7401 Quick-RNA MicroPrep Kit Zymo Research R1051 Quick-RNA MiniPrep Kit Zymo Research R1054 Ethyl alcohol, pure Sigma-Aldrich (continued) E7023 qScript cDNA SuperMix Quanta Biosciences 95,048 OTHER RNase Away Thermo Fisher Scientific 14 375 35 Noyes Spring Scissors - Angled Fine Science Tools 15,013 12

Techniques: Staining, Infection

Journal: EBioMedicine

Article Title: AI-guided discovery of the invariant host response to viral pandemics.

doi: 10.1016/j.ebiom.2021.103390

Figure Lengend Snippet: Fig. 8. Validation of ViP signature-guided therapeutic goals. (a-c) The 166-gene ViP signature-was used to classify liver biopsies from HCV-infected patients treated or not with directly acting anti-viral agents. ROC-AUC values are shown below each bar plot unless otherwise stated. (d) 166-gene ViP signature-based classification of blood samples from HIV- infected patients treated with anti-retroviral therapy (ART). (e) The compound EIDD-2801 (MK-4482; 500 mg/kg) or vehicle (Veh) was administered at indicated doses to Golden Syrian hamsters 4 h prior to intranasal infection with SARS-CoV-2. Hamsters were sacrificed on day 5 and lungs we analyzed by RNA sequencing. (f) Bar (top) and violin (bottom) plots using the ViP (left) or sViP (right) signature-based classification of lung samples from hamsters in E and uninfected controls. (g) Schematic showing the experimental design for validating the ViP signatures as useful tools to assess therapeutic efficacy. Uninf, uninfected; Den3 and Anti-CoV-2 indicate SARS-CoV-2 challenged groups that received either a control mAb or the clone CC12.2 of anti-CoV-2 IgG, respectively. (h) Bar (top) and violin (bottom) plots display the 166- and 20-gene ViP signatures in the uninfected and the SARS- CoV-2 challenged groups, treated with control or anti-CoV-2 IgG. (i-k) Lungs harvested from the 3 groups of hamsters were analyzed by H&E and IHC. Representative images are shown in I. Mag = 10X. Bar graphs in J display the abundance of cellularity and infiltrates in the lungs of the 3 groups, as determined by ImageJ. Violin plots in K display the intensity of staining for IL15RA (top) and IL15 (bottom), as determined by IHC profiler.

Article Snippet: MATERIALS & REAGENTS ANTIBODIES USED FOR IMMUNOCYTOCHEMISTRY Name Manufacturer Catalog number Dilution factor IL15 (E-4) Santa Cruz sc-8437 1:10 IL15RA Proteintech 16,744 1-AP 1:200 Goat anti-rabbit Vector Laboratories, Burlingame, USA MP-7401 1:500 Goat anti-mouse Vector Laboratories, Burlingame, USA MP-7402 1:500 INSTRUMENTS Leica DMI4000B (Automated Inverted Microscope) Leica Microsystems DMI4000B Power Pressure Cooker XL Tristar Products FisherbrandTM 150 Handheld Homogenizer Fisher Scientific (continued) 15,340,168 SOFTWARE ImageJ https://imagej.nih.gov/ij/index.html GraphPad Prism https://www.graphpad.com/scientific-software/prism/ KITS, ENZYMES, CHEMICALS, AND REAGENTS ELISA MAX Deluxe set Biolegend 435,104 V-PLEX sandwich immunoassays Mesoscale Discovery (MSD) K151A9H-1 Zinc Formalin Fisher Scientific 23 313,096 Xylene VWR XX0060 4 Hematoxylin Sigma-Aldrich Inc MHS1 Ethanol Koptec UN1170 Sodium Citrate Sigma-Aldrich W302600 DAB (10X) Vector Laboratories, Burlingame, USA SK-4105 Hematoxylin Sigma-Aldrich Inc. MO, USA MHS1 Stable Peroxidase substrate buffer (10x) Thermo Fisher 34,062 1:10 3% Hydrogen Peroxide Target 245 07 3628 Horse Serum Vector Labs 30,022 Paraformaldehyde 16% Solution, EM Grade Electron Microscopy Sciences 15,710 100% Methanol Supelco MX0485 Glycine Fisher Scientific BP381 5 Bovine Serum Albumin Sigma-Aldrich A9647 100G Triton-X 100 Sigma-Aldrich X100 500ML Prolong Glass Invitrogen P36984 Nail Polish (Rapid Dry) Electron Microscopy Sciences 72,180 Gill Modified Hematoxylin (Solution II) Millipore Sigma 65,066 85 Goat serum Vector Laboratories MP-7401 Quick-RNA MicroPrep Kit Zymo Research R1051 Quick-RNA MiniPrep Kit Zymo Research R1054 Ethyl alcohol, pure Sigma-Aldrich (continued) E7023 qScript cDNA SuperMix Quanta Biosciences 95,048 OTHER RNase Away Thermo Fisher Scientific 14 375 35 Noyes Spring Scissors - Angled Fine Science Tools 15,013 12

Techniques: Biomarker Discovery, Infection, Retroviral, RNA Sequencing, Control, Staining