Fig 2: LAYN promotes CHOL cell proliferation, migration, invasion, and EMT through the TLN1/ITGB1 axis. A. STRING database for predicting proteins that may interact with LAYN. B. Expression box diagram of TLN1. *p < 0.01. C. Correlation analysis between LAYN and TLN1. D. Co-IP assay for validating the interaction between LAYN and TLN1. E. WB assay for detecting TLN1 expression. F. CCK-8 assay for detecting cell proliferation. G. Transwell assay for detecting cell migration. H. Transwell assay for detecting cell invasion. Scale bar = 100 μm. I. IF assays for detecting Snail expression. Scale bar = 50 μm. J. WB assay for detecting E-cadherin, N-cadherin, and Snail expression. K. WB assay for detecting LAYN, TLN1, and p-TLN1 expression. L. WB assay for detecting ITGB1, c-Jun, ERK, p38, p-c-Jun, p-ERK and p-p38 expression. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. n = 3. Non-adjusted images of blots are shown in Supplementary Fig. 3.

Fig 3: CREB1 may be an upstream target of LAYN. A. Expression box diagram of CREB1. *p < 0.01. B. Correlation analysis of CREB1 with LAYN and TLN1. C. JASPAR database for prediction of CREB1 motifs and promoter sequences for possible targeting. D. ChIP assay for validating the binding of CREB1 to the LAYN promoter. *p < 0.05, **p < 0.01. n = 3.