Kinetics of osmotic stress regulate a cell fate switch of cell survival-Step-ID-91719-Jurkat cells exposed to hypertonicity

To understand the molecular changes of signaling and markers of cell state when cells are exposed to hypertonic stress, we measured time courses for the below markers: comb step combined comb step FSC_H comb step FSC_W comb step FSC_A comb step SSC_H comb step SSC_W comb step SSC_A combaba step SC_MKP1_FITC combaba step BD_pS6_A647 combaba step SC_NFAT5_A594 combabb step CS_aCaspase8_PE combabb step BD_aCaspase3_A647 combabc step BD_aCaspase3_PE combabc step BD_cPARP_A647 combabd step CS_pJNK_A647 combabd step CS_pMKK4_PE combabe step BD_IFNy_A700 combabe step BD_IL1b_PE combabf step BD_pH2AX_A555 combabf step BD_Ki67_A700 combabg step BD_pH2AX_A555 combabg step BD_FAS_PECy5 combabh step BD_pCREB_A647 combabh step BD_pmTOR_PE combabk step CS_p27kip1_PE combabk step CS_pHsp27_A647 combabi step CS_CHOP_A647 combabi step CS_LC3_PE combabj step CS_aCaspase9_PE combabj step BD_pNfkb_A647 combabp step CS_PP2AC_PE combabp step CS_pp38_A700 combabl step CS_p38_PE combabl step SC_Caspase12_A647 combabm step CS_pBad_PE combabm step BD_pMK2_A647 combabn step BD_FAS_PECy5 combabn step BD_FASL_PE combabo step BD_Bcl2_PE combabo step CS_BclXL_A647 combabv step CS_pp38_A594 combabv step SC_pASK_A680 combabq step SC_NOXA_A594 combabq step SC_ppp2ac_A680 combabr step RD_NLRP3_A700 combabr step CS_pATF2_PE combabs step SC_Bax_A647 combabs step CS_pIKKab_PE combabt step CS_pStat3_A647 combabt step CS_LC3b_PE combabu step SC_Bid_A647 combabu step BD_cPARP_PE combabw step BD_TNF_A700 combabw step CS_pcJun_PE combabx step BD_pBcl2_PE combabx step SC_HSP70_A680 combaby step CS_pAMPK_A647 combaby step SC_Ub_A594 combabz step CS_pATG14_A647 combabz step SC_Ub_A594 combabz2 step CS_pp38_A647 combabz2 step SC_Ub_A594 combabz3 step CS_antirabbit_igg_A647 combabz3 step SC_Ub_A594
Conclusion:
Conclusions are described in: https://www.biorxiv.org/content/10.1101/2020.07.10.197871v1
Notes:
timepoints 0,2,5,10,15,20,30,60,120,180,240 300 min after addition of 300 mosmol/l NaCl are barcoded by 12 barcodes using Pacific-Blue (dilution: high, mid, low) and Pacific-Orange ((dilution: high, midhigh, midlow, low) following established protocols for fluorescent cell barcoding for flow cytometry: timepoints are barcoded as follows: PO high + PB high=0, PO midhigh + PB high=2, PO midlow + PB high=5, PO low + PB high=10, PO high + PB mid=15, PO midhigh + PB mid=20, PO midlow + PB mid=30, PO low + PB mid=60, PO high + PB low=120, PO midhigh + PB low=180, PO midlow + PB low=240, PO low + PB low=300