Journal: Clinical and Translational Medicine
Article Title: ZSH‐2208: A novel retinoid with potent anti‐tumour effects on ESCC stem cells via RARγ–TNFAIP3 axis
doi: 10.1002/ctm2.70148
Figure Lengend Snippet: ZSH‐2208 inhibits the transcription of TNFAIP3 through reducing the expression of protein of RARγ. (A) Pattern diagram of molecular docking simulation of ZSH‐2208 to RARγ protein. (B–D) Effect of ZSH‐2208 on RARγ protein and gene expression levels in ESCC‐TRCs. (E) Heatmap of the significantly upper (lower)—modulated differentially expressed gene (DEGs) Top50 after ZSH‐2208 treatment. (F) Combined analysis of sequencing by RNA‐seq and ChIP‐seq identified TNFAIP3, a key effector molecule for ZSH‐2208. (G) ZSH‐2208 treatment significantly reduced the binding peak of RARγ protein in the promoter region of TNFAIP3 gene. (H) Three possible binding sites for the RARγ protein on the TNFAIP3 gene promoter. (I) Effect of ZSH‐2208 on TNFAIP3 and TNIP1 at protein level in ESCC‐TRCs. (J) Immunofluorescence staining of TNFAIP3 protein was performed in subcutaneous tumour tissues of nude mice to observe the effect of ZSH‐2208 on TNFAIP3 protein expression. DOX, doxorubicin; red, TNFAIP3: blue. (K–M) Effect of CD2665 on TNFAIP3 and TNIP1 at gene and protein level in ESCC‐TRCs (* p < .05, ** p < .01, *** p < .001, **** p < .0001; ns, not significant).
Article Snippet: Additionally, we treated ESCC‐TRCs with CD2665 (MCE, HY‐107437, USA), a selective antagonist of RARγ, and observed a reduction in the expression levels of both TNFAIP3 and TNIP1 at the gene level.
Techniques: Expressing, Sequencing, RNA Sequencing Assay, ChIP-sequencing, Binding Assay, Immunofluorescence, Staining
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