Fig. 6: ZBP1 deficiency impairs CAF-mediated CCL7 secretion and attenuates tumor cell malignancy via the CCL7–CCR1 axis.

A qRT-PCR validation of ZBP1 mRNA deficiency in CAFs. B Western blot analysis was conducted to confirm the deficiency of ZBP1 in CAFs. C qRT-PCR analysis of CCL7 mRNA expression in CAFs under control conditions and following ZBP1 deficiency. D ELISA was utilized to measure the levels of CCL7 in the supernatant of CAFs before and after ZBP1 deficiency. E GSVA heatmap was generated to illustrate the activities of hallmark pathway in CAFs from WT and Zbp1^−/− groups. F Western blot analysis was performed to evaluate p65/IκBα signaling to assess NF-κB pathway activity in CAFs. Samples include sgNS controls, sgNS treated with BAY11-7085 (NF-κB inhibitor), sgZbp1, and sgZbp1 treated with TNF-α (an NF-κB activator). G qRT-PCR analysis was conducted to assess Ccl7 mRNA expression in CAFs. H ELISA analysis was performed to determine the levels of secreted CCL7 levels in CAF supernatants. I Western blot analysis was conducted to assess CCR1 expression in MOC1 cells co-cultured with differently treated CAFs (sgNS or sgZbp1). J A schematic diagram illustrates the Transwell assay employed to evaluate the impact of various co-culture conditions on the migration and invasion of MOC1 cells. K The migration and invasion capabilities of MOC1 cells were analyzed after 48 h under different co-culture conditions, accompanied by statistical analysis. L A schematic diagram depicts the EdU assay used to evaluate MOC1 cell proliferation under varying co-culture conditions. M The proliferation of MOC1 cells was assessed after 24 h under different co-culture conditions, with accompanying statistical analysis.