Fig. 6

CREB3L1 is involved in remodeling the stromal microenvironment of ATC. A Single-cell RNA-sequencing was used to examine the interaction between ATC cell subsets and cells in the tumor microenvironment. B The sphere formation assay was used to detect the growth of 8505C cells mixed with or without CAFs after CREB3L1 knockdown. ***P < 0.001 CAF + NC versus CAF + CREB3L1-KD. C Schematic diagram of the experimental procedure. Equal numbers of DiI-labeled 8505C cells (red) and DiO-labeled CAFs (green) were mixed and implanted into the perivitelline space of each zebrafish. D The zebrafish xenograft model was employed to evaluate the CAF-mediated metastasis of 8505C cells after CREB3L1 knockdown. E Flow cytometry analysis of α-SMA-positive fibroblasts, after co-culture with 8505C cells. *P < 0.05, **P < 0.01 versus the respective CAF + medium, CAF + NC, or CAF + CREB3L1-KD. F Flow cytometry analysis of α-SMA positive fibroblasts in the sphere derived from the mixture of 8505C and CAFs. G Flow cytometry analysis of α-SMA-positive fibroblasts in the lung tissues of mice with ATC pulmonary metastasis. *P < 0.05 NC versus CREB3L1-KD. H Counterstaining of Ki67 and α-SMA with antibodies that specifically reacted with the human (Ki67) and mouse (α-SMA) antigen, respectively. Data are presented as the mean ± SD