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Fig. 3 | Journal of Experimental & Clinical Cancer Research

Fig. 3

From: PD-1 blockade delays tumor growth by inhibiting an intrinsic SHP2/Ras/MAPK signalling in thyroid cancer cells

Fig. 3

Signalling pathways downstream PD-1 activation/overexpression. a. Activation of BRAF, MEK1/2 and MAPK (p44/p42) in 8505c and TPC-1 cells, transfected with pFLAG PD-1 or the relative empty vector (pFLAG), assessed by western blot for their phosphorylated forms. A representative experiment is shown. b. Activation of BRAF, MEK1/2 and MAPK in 8505c and TPC-1 cells, treated or not with sPD-L1 (1 μg/ml - 30 min), assessed by western blot for their phosphorylated forms. A representative experiment is shown. c. Activation of BRAF, MEK1/2 and MAPK in 8505c and TPC-1 cells, treated with siPD1 or siCTR (100 nM - 48 h) or with Nivolumab or IgG4 (10 μg/ml – 15 and 30 min), assessed by western blot for their phosphorylated forms. A representative experiment is shown. d. Pull-down assay with the GST-RAF1-Ras Binding Domain (RBD) of 8505c and TPC-1 cells transiently transfected with pFLAG + pCEFL, pFLAG PD-1 + pCEFL, pFLAG + pCEFL H-Ras AU5, or pFLAG PD-1 + pCEFL H-Ras AU5. A representative pull-down is shown, together with the mean densitometric analysis ± SD of 5 independent assays. * P < 0.05 compared to the relative control

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