Fig 2: PGLS regulated the proliferation and apoptosis of HCC. (A) Transfection with the three siRNAs could effectively knock down the expression of PGLS on Huh7. (B) Quantitative analysis of Western blot in (A). (C) The NADP+/NADPH ratio on Huh7. (D) The percentage of ROS on Huh7. (E) The apoptosis rate on Huh7. (F) Fluorescence micrographs of Huh7 after staining with calcein-AM, PI, and DAPI. (G) Proliferation curve of Huh7 transfected by siRNA-1, siRNA-2, and siRNA-3. (H) Representative field of CFUs, formed by Huh7 treated with siRNA-1, siRNA-2, and siRNA-3. (I) CFU number per field on Huh7. Data show individual values and mean ± SD. (B–E,G,I) Unpaired two-tailed Student t-tests, assessed statistical significance, ***p < 0.001.

Fig 3: PGLS regulated the migration and invasion of HCC. (A) Representative images of CFUs in migration assay. (B) CFU clone number per field on Huh7 in migration assay. Invaded cells from five representative fields were counted. (C,D) The number of invasive cells significantly decreased after the treatment of these three siRNAs, respectively (n = 5). (E) Wound healing in Huh7 cells treated with siRNA-1,siRNA-2, and siRNA-3. The lines indicated the edge of wound at 0 and 24 h. (F) Migration rate was analyzed and expressed as the area of cells migrating from the original wounds. Data show individual values and mean ± SD. (B,D,F) Unpaired two-tailed Student t-tests, assessed statistical significance, ***p < 0.001.

Fig 4: PGLS pathway led to significant activation of PPP in HCC. (A) The representative FACS plot of PGLShigh and PGLSlow HCC cells. (B,C) Western blot of the PGLS and CK18 in PGLShigh and PGLSlow cells from human HCC patients. ß-Actin is a loading control. (D) The relative expression of HCC marker genes in PGLShigh and PGLSlow cells (n = 3 replicates). (E) Glycometabolism diagram. (F) The NADP+/NADPH ratio in PGLShigh and PGLSlow cells (n = 5 replicates). (G) The percentage of ROS in PGLShigh and PGLSlow cells (n = 5 replicates). (H) The representative FACS plot of PGLShigh and PGLSlow cells for apoptosis analysis. (I) The apoptosis rate of PGLShigh and PGLSlow cells. (J) The expression profile of genes associated with apoptosis pathway between PGLShigh and PGLSlow cells. (K) KEGG analysis of PGLShigh and PGLSlow cells. Data show individual values and mean ± SD. (C,D,F,G,I) Unpaired two-tailed Student t-tests, assessed statistical significance, *p < 0.05, **p < 0.01, ***p < 0.001.