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Fig. 4 | Journal of Experimental & Clinical Cancer Research

Fig. 4

From: Argininosuccinate synthase 1 suppresses tumor progression through activation of PERK/eIF2α/ATF4/CHOP axis in hepatocellular carcinoma

Fig. 4

ASS1 is upregulated through ER stress response in HCC spheroids (a) Microarray analysis of ASS1-overexpressing HCC spheroids implicated four genes for further study (ASNS, ATF3, CHOP, and HSPA1A). (b) Expression of ER stress response related proteins; GRP78, XBP1us, CHOP and ATF3 in monolayers (2D) or spheroids (3D) of HCC cell lines including SNU475, SNU449 and Huh7 cells. (c-d) ASS1 mRNA levels analysis in Huh7 cells and SNU475 cells with treatment of ER stress inducers; TG and TM by qRT-PCR. (e-f) ASS1 promoter activity by luciferase assay in Huh7 cells with treatment of TG and TM. ASS1 expression levels in Huh7 and SNU475 cells with treatment of (g) TG and (h) TM. (i) Immunofluorescence (IF) images showing ASS1 (green), ER (red), and nuclei (blue) in Huh7 cells with TG and TM treatment. *P < 0.05, **P < 0.01 and ***P < 0.001 compared to control group. At least triplicate analysis in each set of experiment. TG; thapsigargin, TM; tunicamycin

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