Fig. 1: GRP78-induced macrophages promote CRC chemoresistance and enhance tumor cell stemness.

A The tSNE clustering plot for single cells. B The tSNE clustering plot for macrophage subsets. C Box plot of enrichment of M0, M1, and M2 macrophages in high and low-risk groups for 5-FU resistance. D Diagram of a cellular communication network. The size of the dots in the diagram represents the relative number of cells, and the thickness of the connecting lines represents the strength of intercellular communication. E The effects of M0-CM and GRP78-CM on the viability of HCT-8 and DLD1 cells were detected by MTT and IC₅₀ values were calculated. F The effects of M0-CM and GRP78-CM on 5-FU-induced apoptosis in HCT-8 and DLD1 cells were detected by flow cytometry. G Statistical plots of apoptosis rates of HCT-8 and DLD1 cells for (F). H The effects of M0-CM and GRP78-CM on the expression of pro-apoptotic proteins cleaved-PARP and cleaved-caspase 3 and anti-apoptotic protein Bcl-2 in the presence of 5-FU by western blot. I The effects of M0-CM and GRP78-CM on the expression of HCT-8 and DLD1 cell stemness markers CD133, CD44 and ALDH1A1 proteins by western blot. J The effects of M0-CM and GRP78-CM on the expression of stemness markers CD133, CD44 and ALDH1A1 mRNA in HCT-8 and DLD1 cells as detected by qRT-PCR. Data are expressed as SD ± mean, n = 3. *p < 0.05, **p < 0.01, ***p < 0.001.