Extended Data Fig. 7: DDX5 mediates the change of splicing events and IL-36R/sIL-36R expression.
a, Analysis of different splicing events in IL-36γ-stimulated wild-type and DDX5–/– HaCaT cells (n = 3), MC903-treated Ddx5fl/fl(n = 3) and Ddx5∆KC (n = 4) mice or IMQ-treated Ddx5fl/fl and Ddx5∆KC mice (n = 3). SE, skipped exon; MXE, mutually exclusive exon; A5SS, alternative 5’ splice site; A3SS, alternative 3’ splice site; RI, retained intron. b, DNA-PAGE of splicing variants of IL4R, TSLPR, IL7R, IL17RA, IL17RC, CD6, FGFR2 and IL20RB in wild-type and DDX5–/– HaCaT cells. Data represent three independent experiments. c-d, Agarose gel analysis of IL36R transcripts amplified by PCR in NHEKs (c) or primary murine keratinocytes (d). e-f, Schematic diagrams of the structure of human (e) or murine (f) IL-36R and sIL-36R. g, Immunoblot of full-length IL-36R or sIL-36R by the antibody against Flag (left panel) or the specific antibody against murine sIL-36R (Middle panel) or the specific antibody against human sIL-36R (Right panel). h, Immunoblot of sIL-36R in cell cultures or cell lysates from HaCaT cells in which Flag-tagged human sIL-36R was overexpressed. Data represent three independent experiments. i, DNA-PAGE of IL36R and sIL36R transcripts in wild-type and DDX5–/– HaCaT cells. j, Silencing efficiency of indicated genes in NHEKs (n = 3). k, DNA-PAGE of Il36R and sIl36R transcripts in primary murine keratinocytes isolated from Ddx5fl/fl and Ddx5∆KC newborn mice. Data represent three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001. n.s. no significance. P values were analyzed by two-way ANOVA (j). Data are presented as mean ± s.e.m.
