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. 2016 May 4;7:11379. doi: 10.1038/ncomms11379

Figure 7. The HCV core altered the ER distribution and induced ER stress in SPP/TRC8DKO Huh7 cells.

Figure 7

(a) SPP/TRC8DKO Huh7 cells (bottom) and those stably expressing SPP-HA (top) were transfected with pCAG OSF-HCV core and pCMV DsRed-ER and incubated for 48 h. FLAG-HCV core (green) and nuclei (blue) were stained with anti-FLAG antibody and DAPI, respectively. Scale bar, 40 μm. Images are representative of two independent experiments. (b) The luciferase activity was determined in cells transfected with a reporter plasmid containing spliced XBP1 (ERAI) and pRL-SV40 and treated with 1 μM of thapsigargin (TG) or tunicamycin (TU) 24 h post transfection. The data represent the mean±s.d. of two independent experiments. (c) The luciferase activity was determined in cells transfected with plasmids encoding the HCV core, ERAI and pRL-SV40 at 24 h post transfection. Significant enhancement (*P<0.05) of XBP1s production was observed with HCV core expression only in SPP/TRC8DKO Huh7 cells. The data represent the mean±s.d. of two independent experiments performed with a cell line representative of Huh7 cells. (d) RNA was extracted from cells transfected with an HCV core protein expression plasmid of at 24 h post transfection, and binding of immunoglobulin protein (BIP; left) and CCAAT/enhancer-binding protein homologous protein (CHOP; right) expression levels were quantified by qPCR. The data represent the mean±s.d. of two independent experiments performed with a cell line representative of Huh7 cells. (e,f) The luciferase activity was measured in cells transfected with plasmids encoding the EHcV core (e) or JEV core (f), ERAI and pRL-SV40 at 24 h post transfection. The data represent the mean±s.d. of two independent experiments performed with a cell line representative of Huh7 cells. The significance of the differences was determined using Student's t-test.