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. 2018 Sep 24;9(10):971. doi: 10.1038/s41419-018-1039-2

Fig. 4. GRP75 inhibition attenuates the degradation of proto-Dbl.

Fig. 4

a GRP75 knockdown stable cell lines (GRP75-KD: G1, G2, G3) were produced from SKOV-3 cells (WT) by the CRISPR–CAS9 system. pLenti-CRISPR v2 plasmid transfected SKOV-3 cells was set as the control (NC). The protein levels of endogenous proto-Dbl, onco-Dbl and GRP75 were determined by Western blot using the indicated Abs. c SKOV-3 cells were treated with/without CHX (100 µg/ml) for 12 h to inhibit de novo protein synthesis, and the GRP75 inhibitor MKT077 was added either in a time-dependent or in a concentration-dependent manner. The protein levels of endogenous proto- and onco-Dbl were analyzed by Western blot. e GRP75-KD stable cell lines (G2, G3) were pre-treated with the UPP inhibitor MG132 (50 μM, 6 h) before harvest. DMSO treatment was used as the control. The protein level changes of proto- and onco-Dbl after MG132 treatment was analyzed by Western blot. Representative blot results from three independent experiments are shown in a, c, e, and corresponding quantitation data (determined similarly as above) are shown in b, d, f. Statistically significant differences compared with corresponding NC groups are shown: **P < 0.01