Fig 1: Integration of single-cell expression data and PERK pathway inhibition demonstrates ER-stress–dependent regulation of GP73 in hepatocellular carcinoma. (A) t-SNE plots depicting clustering of malignant (left) and non-malignant (right) cells, color-coded by patient (malignant) or cell type (stroma). (B) Expression of GOLM1 and EIF2αK3 mapped onto t-SNE plots, derived from Sequential NCI-CLARITY (GSE229772) and analysed using the Single-Cell Atlas in liver cancer (scAtlasLC) (C) Kaplan-Meier survival curves of HCC patients with low and high expression of GP73, derived from the Cancer Genome Atlas (TCGA). (D-E) Immunohistochemistry images of EIF2αK3 and GOLM1 expression levels in HCC biopsies, derived from the Human Protein Atlas, proteinatlas.org. (F) Representative GP73-antibody-stained immunohistochemistry images from mouse livers. Scale bar: 200 μm. (G) Quantification of the GP73-positive area. (H-J) Quantification of GP73 secretion in HepG2, Huh7, and SNU449 cells treated with thapsigargin alone or in combination with AMG-PERK. Bars represent mean with ± SD. N = 7 mice per group for GP73 IHC.
Fig 2: PERK inhibition reduces GP73-associated ER-stress and fibrotic responses in hepatic stellate cells. (A) mRNA expression of αSMA, CTGF, PERK, and DDIT3 in LX-2 cells treated with tumor-conditioned medium (T.M.) derived from Huh7 cells (B) Representative αSMA immunofluorescence images of LX-2 cells treated with T.M., T.M. + thapsigargin (Thapsi), T.M. + Thapsi + AMG-PERK, T.M. + Thapsi + a blocking antibody against the extracellular binding site of the surface receptor GRP78 (C) Quantification of αSMA staining normalized to nuclear staining (DAPI). (D) Representative ER-Tracker staining under the above-mentioned conditions. (E) Quantification of ER-Tracker signal normalized to nuclear staining (DAPI). (F) mRNA expression of αSMA, CTGF, PERK, and DDIT3 in LX2 cells treated with recombinant GP73. (G-J) Representative images and quantification of αSMA and ER-Tracker staining in LX-2 cells treated with GP73 alone or in combination with a blocking antibody against the extracellular binding site of the surface receptor GRP78. (K-N) qPCR analysis of αSMA, CTGF, PERK, and DDIT3 across control, TM, GP73, TM + GRP78 blockade and GP73 + GRP78 blockade. (O) Schematic representation of a proposed model (created with BioRender.com). Bars represent mean with ± SD. N = 3 biological replicates for qPCR; N = 4 biological replicates for staining.
Supplier Page from Fine Biotech Co., Ltd. for Recombinant Human GOLM1