Fig 2: Negative correlations between SMYD3 (A) or SUV39H2 (B) and chemokines or APM molecules(A) SMYD3 mRNA was negatively correlated with mRNA levels of CXCL9, CXCL10 and CXCL11 chemokines, and with mRNA levels of TAPBP, TAP1 and TAP2 using the TCGA database for SCCHN. (B) SUV39H2 mRNA was negatively correlated with mRNA levels of CXCL9, CXCL10 and CXCL11 chemokines, and with mRNA levels of HLA-A, B2M and TAP1 using the TCGA database for SCCHN. Pearson's correlation coefficients r were calculated and significance was defined as P < 0.05. All shown results were statistically significant.

Fig 3: Functional interaction between SMYD3, AMPK, and mTOR. Co-immunoprecipitation of endogenous SMYD3, AMPK, or mTOR in HCT-116 and MDA-MB-231 cells treated or not with NCS (5 nM) for 24 h. Anti-IgGs were used as a negative control.

Fig 4: Effects of NCS on the AMPK/mTOR pathways after pre-treatment with EM127. (a–c) Immunoblotting showing the levels of proteins involved in the AMPK and mTOR pathways in HCT-116 (a), AGS (b) and MDA-MB-231 (c) cells pre-treated with the SMYD3 inhibitor EM127 (5 μM) for 24 h and then treated with NCS (5 nM) for 24 h. VINCULIN was used as a loading control.

Fig 5: SMYD3 is required for regulating EMT, and depletion of SMYD3 expression attenuates sorafenib-resistant HCC cell migration, invasion, tumorigenicity, and self-renewal properties(A and B) SMYD3 expression in sorafenib-resistant cells after stable SMYD3 knockdown was assessed by western blot (A) and RT-PCR (B). Data are represented as mean ± SD.(C and D) The expression of EMT markers in SMYD3-depleted sorafenib-resistant Huh7 and SMMC-7721 cells was detected by RT-PCR (C) and western blot (D). Data are represented as mean ± SD.(E and F) Transwell assays (E) and colony and spheroid formation showed the migration and invasion abilities (E) and the tumorigenicity in vitro and self-renewal abilities (F) of SMYD3-depleted sorafenib-resistant cells. Data are represented as mean ± SD.