Growth Protocol:	-
Treatment Protocol:	CD16High and CD16Int neutrophils were sorted by FACSAria III from three severe COVID-19 patients and NDN were purified from healthy donor peripheral blood.
Extract Protocol:	RNA was extracted using Qiagen's RNAeasy Plus Microkit.
Library Construction Protocol:	Library Preparation: Libraries were prepared using the Universal Plus mRNA-Seq with (NuGEN Cat# 0520-24). a. Poly(A) Selection: 8-30 ng of RNA samples (in a volume of 50 μl) were mixed with Oligo (dT) Beads and incubated at 65oC for 5 minutes, mixed, then at room temperature for 10 mins. The supernatant was discarded and the beads were washed with wash buffer. Captured polyadenylated RNA was eluted using Elution buffer at 80°C for 2 min. mRNA is further purified in a second bead clean-up. b. RNA Fragmentation: Beads were mixed with 20 μl Fragmentation Buffer and incubated for 8 minutes at 94oC. 20 μl of supernatant was removed from the beads and proceeded immediately to First Strand cDNA Synthesis. c. First Strand cDNA Synthesis: 5 μl of First Strand Master Mix was added to each sample and placed on a thermal cycler to run preprogrammed thermal conditions. d. Second Strand cDNA Synthesis: 50 μl Second Strand Master Mix was added to each sample, mixed well and incubated in a thermal cycler at 16oC for one hour. e. cDNA Purification: cDNA was purified using Agencourt AMPure XP Beads (1.8 volumes), eluted in 10 μl of H2O, and stored at -20oC. f. End Repair: End Repair Mix was added to the purified samples from step e and the samples were incubated in a preprogrammed thermal cycler. g. Adaptor Ligation: Samples were mixed with 3 μl adaptor mix, then 12 μl Ligation Mix, and incubated in a thermal cycler at 25oC for 30 minutes. Sample and Barcode Information: Samples were barcoded with NuGEN Unique Dual Indexes. Strand Selection: 70 μl of Strand Selection Master Mix was added to 30 μl of each sample, incubated in a pre-heated thermal cycler at 72oC for 10 minutes. Strand Selection Purification: Strand selected cDNA was purified using Agencourt AMPure XP Beads (0.8 volumes), eluted in 15 μl of H2O, and stored at - 20oC. j. Library Amplification: 76.5 μl of Library Amplification Master Mix was added to 13.5 μl of each sample and incubated in a preprogrammed thermal cycler, for 13- 16 cycles, which was pre-optimized with qPCR. k. Amplified Library Purification: Amplified libraries were purified using 90 μl Agencourt AMPure XP Beads (1.0 volumes). 25 μl of eluted libraries were collected and stored at -20oC. l. Validate Library: The concentration of libraries were measured by Qubit dsDNA HS Assay Kit (Invitrogen Q32851). Libraries were diluted and normalized to the optimal range for Agilent Bioanalyzer analysis using the DNA High Sensitivity Kit (Agilent Technologies, Cat# 5067-4626). m. Normalize and Pool Libraries: Same amount of libraries were pooled based on the molar concentration from Bioanalyzer measurements. Library Denaturing and Diluting for MiSeq Nano 300: Pooled library was run on MiSeq to test quantity and quality, using the MiSeq Reagent Nano Kit V2 300 cycles (Illumina, Cat. No. MS-103-1001). Library and PhiX control (Illumina, Cat. No. FC-110-3001) were denatured and diluted using the standard normalization method following manufacturer's directions, to a final concentration of 6pM. Equal volume of library and PhiX were combined and sequenced on Illumina MiSeq. (https://basespace.illumina.com/run/196126950/JY_Xiaoling07072020_test) 3. Library Denaturing and Diluting for Nextseq 500: Library and PhiX were denatured and diluted using the standard normalization method following manufacturer's directions. The loading of library pool was 1.3 ml at 1.8 pM, with 1% PhIX spike in. Sequencing Run: Sequencing was performed on the University of Louisville Brown Cancer Center Genomics Core Illumina NextSeq 500 using the NextSeq 500/550 75 cycle High Output Kit v2.5 (20024906).

Molecule Type:	polyA(+) RNA
Library Source:
Library Layout:	SINGLE
Library Strand:	Reverse
Platform:	ILLUMINA
Instrument Model:	Illumina NextSeq 500
Strand-Specific:	specific

Data Resource	GEN Sample ID	GEN Dataset ID	Project ID	BioProject ID	Sample ID	Sample Name	BioSample ID	Sample Accession	Experiment Accession	Release Date	Submission Date	Update Date	Species	Race	Ethnicity	Age	Age Unit	Gender	Source Name	Tissue	Cell Type	Cell Subtype	Cell Line	Disease	Disease State	Development Stage	Mutation	Phenotype	Case Detail	Control Detail	Growth Protocol	Treatment Protocol	Extract Protocol	Library Construction Protocol	Molecule Type	Library Layout	Strand-Specific	Library Strand	Spike-In	Strategy	Platform	Instrument Model	Cell Number	Reads Number	Gbases	AvgSpotLen1	AvgSpotLen2	Uniq Mapping Rate	Multiple Mapping Rate	Coverage Rate