Dataset Information

Differentially expressed proteins in Human Primary Keratinocytes upon IL-9 stimulation

ABSTRACT: HPKs from 3-5 healthy individual donors were pre-stimulated with TGFb (10ng/ml) plus IL-4 (20ng/ml) to upregulated the IL-9R (Das et al., 2019). Next day, HPKs were stimulated with IL-9 (50ng/ml) or in combination with IFNg (10ng/ml) for 24 hours at 37 degree C with 5% CO2. Sample preparation for LC-MS/MS was performed as described previously with few modifications. HPKs were lysed in TRIzol reagent (Invitrogen, Carlsbad, CA) and proteins were extracted as per the guidelines by the manufacturer. The instruments were set to MS1 resolution of 70000 and MS2 resolution of 17500. The MS spectra were analysed using the Thermo-scientific mass informatics platform Proteome discoverer version 2.2. Workflows for discovery proteomics were used with both Mascot and SequestHT as search engines. For label-free quantification, the standard LFQ workflows by Thermo were used. The samples were normalized with respect to the peak-area based total peptide amount. The permitted retention time shift (RT) was set to 2 min. Fragment mass tolerance was set to 0.02 Da and the precursor mass tolerance to 2ppm. The false discovery rate (FDR) was set to 0.01 (Strict). Importantly, only unique peptides were considered for peak-area based quantification. For statistical analysis of identified proteins, null hypothesis testing was performed using ANOVA (background based).

INSTRUMENT(S): Q Exactive Plus

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Rahul Purwar

PROVIDER: MSV000086776 | MassIVE | Thu Jan 28 03:10:00 GMT 2021

REPOSITORIES: MassIVE

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Publications

Multiomics Analysis and Systems Biology Integration Identifies the Roles of IL-9 in Keratinocyte Metabolic Reprogramming.

Marathe Soumitra S Dhamija Bhavuk B Kumar Sushant S Jain Nikita N Ghosh Sarbari S Dharikar Jai Prakash JP Srinivasan Sumana S Das Sreya S Sawant Abhijeet A Desai Saloni S Khan Farhat F Syiemlieh Abigail A Munde Manohar M Nayak Chitra C Gandhi Mayuri M Kumar Ashutosh A Srivastava Sanjeeva S Venkatesh K V KV Barthel Steven R SR Purwar Rahul R

The Journal of investigative dermatology 20210303 8

IL-9‒producing T cells are present in healthy skin as well as in the cutaneous lesions of inflammatory diseases and cancers. However, the roles of IL-9 in human skin during homeostasis and in the pathogenesis of inflammatory disorders remain obscure. In this study, we examined the roles of IL-9 in metabolic reprogramming of human primary keratinocytes (KCs). High-throughput quantitative proteomics revealed that IL-9 signaling in human primary KCs disrupts the electron transport chain by downregu ...[more]

PMID: 33667432

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