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Fig. 1 | Cell & Bioscience

Fig. 1

From: Regulation of platelet-activating factor-mediated interleukin-6 promoter activation by the 48 kDa but not the 45 kDa isoform of protein tyrosine phosphatase non-receptor type 2

Fig. 1

Differential effects of PTPN2 isoforms on PAF-induced IL-6 expression. a HEK-PAFR were transiently co-transfected with the hIL-6-luc, control vector (pcDNA3) or phosphatase constructs a WT and b D182A mutants. Cells were incubated overnight in DMEM-0.2% BSA and stimulated with a, b PAF (100 nM), vehicle or a TNF-α (20 ng/ml) for 6 h and luciferase activity was measured. The data presented are a mean ± SEM of 6–16 independent experiments performed in triplicate. Significance was established with two-way ANOVA with Sidak post-test. *p < 0.05. c iMo-DCs were either not transfected (NT) or transfected with siRNAs: either control (siCTRL) or against isoforms of PTPN2 (siPTPN2 48 kDa or 45 kDa) on day 4 and 5 before being collected on day 7 for experiments. iMo-DCs were starved for 3 h in RPMI 0.2% BSA and stimulated with PAF (1 nM) for 5 h. Cells were lysed in Trizol and RNA was extracted and converted to cDNA. IL-6 mRNA was quantified by real-time PCR. Data are presented as mean ± SEM of mRNA expression calculated by the delta–delta (ΔΔ)Ct method on their respective unstimulated control for 4 independent experiments, using RPL13A and GAPDH as housekeeping genes. Significance was established with two-way ANOVA with Sidak post-test, *p < 0.05

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