Skip to main content
. Author manuscript; available in PMC: 2021 Feb 6.
Published in final edited form as: Mol Cell. 2019 Dec 3;77(3):542–555.e8. doi: 10.1016/j.molcel.2019.11.007

Figure 4: m6A promotes alternative splicing of CIRBP.

Figure 4:

(A) Schematic of CIRBP transcript isoforms with a focus on the alternatively spliced region (ASR). RT-qPCR primer locations are indicated with arrows (FC-RC: control CIRBP amplicon; F-RL: long isoform specific; F-RS: short isoform specific. (B) RT-qPCR analysis of short (S) and long (L) CIRBP RNA isoforms in mock- and virus-infected (48 hpi) Huh7 cells relative to control CIRBP amplicon. (C) RT-qPCR analysis of S and L CIRBP RNA isoforms in mock- and TG-treated (16 h) Huh7 cells. (D) (Left) Representative immunoblot of short (CIRBP-S) and long (CIRBP-L) CIRBP protein isoforms in mock- and virus-infected (48 hpi) Huh7 cells. (Right) Quantification of CIRBP protein isoform expression relative to tubulin. (E) (Left) Representative immunoblot analysis of CIRBP protein isoforms in mock- and TG-treated (500nM, 16 h) Huh7 cells. HSPA5 and GADD34 are positive controls. (Right) Quantification of CIRBP protein isoform expression relative to tubulin. (F) Schematic of WT and m6A-mut CIRBP splicing reporters. RT-qPCR primer locations (Fluc-Rluc: control; F-RL: long isoform specific; F-Rs: short isoform specific) are indicated with arrows. (G) RT-qPCR analysis of CIRBP splicing reporter isoform expression (S and L) relative to control RLuc amplicon in Huh7 cells transfected with WT and m6A-mut constructs. Values are the mean ± SEM of 3 (B, D, E, G) or 5 (C) biological replicates. * p < 0.05, ** p < 0.01, *** p < 0.001 by unpaired Student’s t test. n.s. = not significant. See also Figure S4.