|
Fig. 5
STEAP3-AS1 binds to YTHDF2 to prevent m6A-mediated degradation of STEAP3 mRNA. A The potential binding proteins of STEAP3-AS1 predicted by the AnnoLnc2 database. B m6A modification of STEAP3 mRNA and STEAP3-AS1 in SW480 and DLD-1 cells analyzed by MeRIP. C The interaction between YTHDF2 and STEAP3-AS1 in DLD-1 and SW480 cells as measured by RNA pulldown. Anti-sense was used as a negative control. D RIP analysis of the binding of YTHDF2 to STEAP3 mRNA and STEAP3-AS1 in DLD-1 and SW480 cells. E The binding of YTHDF2 to indicated truncated STEAP3-AS1 in SW480 and DLD-1 cells measured by RNA pulldown. F RNA pulldown analysis of the interaction between YTHDF2 and STEAP3 mRNA in control or STEAP3-AS1-knockdown DLD-1 and SW480 cells. G RIP analysis of the interaction between YTHDF2 and STEAP3 mRNA in DLD-1 and SW480 cells with or without STEAP3-AS1 knockdown. (sh#1, shSTEAP3-AS1 #1). H Relative mRNA level of STEAP3 in control or STEAP3-AS1-knockdown DLD-1 and SW480 cells treated with actinomycin D (2.5 ?M) at indicated time points. (sh#1, shSTEAP3-AS1 #1; sh#2, shSTEAP3-AS1 #2). I Relative STEAP3 mRNA level in control or STEAP3-AS1-overexpressing HCT116 cells treated with actinomycin D (2.5 ?M) at indicated time points. (OE, STEAP3-AS1 overexpression). Data are means ± s.d. and are representative of at least 3 independent experiments. (** P < 0.01 and *** P < 0.001, NS, not significant)