Figure 7.
ELAVL1 is required for TNF/IFNγ-induced cell death. (A and C) HeLa (A) or HCT-116 (C) cells (parental, ELAVL1 KO clones #1 and #2 with different gRNA) were treated with TNF/IFNγ for 48 h (n = 3). Cell viability was determined by ATP levels. One-way ANOVA was performed: ****P < 0.0001. (B and D) The protein expression of ELAVL1 in indicated cells was analyzed by immunoblotting (n = 2). (E and G) HeLa (E) or HCT-116 (G) cells (parental, ELAVL1 KO, ELAVL1 KO stably expressed with WT ELAVL1 or mutated ELAVL1) were treated with TNF/IFNγ for 48 h (n = 3). Cell viability was determined by ATP levels. One-way ANOVA was performed: NS, ****P < 0.0001. (F and H) The protein expression of ELAVL1 in indicated cells was analyzed by immunoblotting (n = 2). (I) The binding interface between ELAVL1 and RNA in the ELAVL1-RNA complex (PDB 4ED5) is shown. Amino acids critical for the RNA binding of ELAVL1 are highlighted. (J) HeLa cells (parental, ELAVL1 KO clones #1 and #2 with different gRNA) were treated with IFNγ for 24 h (n = 3). The mRNA levels of CASP8 were measured by qPCR. One-way ANOVA was performed: ***P = 0.0003, ****P < 0.0001. (K) HeLa cells (parental, CASP8 KO, ELAVL1 KO clones #1 and #2 with different gRNA) were treated with IFNγ for 24 h. The expression of indicated proteins was analyzed by immunoblotting (n = 4). (L and M) HeLa cells (parental and ELAVL1 KO) were treated with actinomycin D for the indicated time. The mRNA levels of CASP8 (L) and CYLD (M) were measured by qPCR (n = 3). The mRNA levels in HeLa cells before the addition of actinomycin D (0 h) were set as 100%. Two-tailed t test was performed: **P = 0.0024 (L); NS (M). (N and O) The mRNA levels of CASP8 (N) and CYLD (O) in HeLa cells (parental, ELAVL1 KO, ELAVL1 KO stably expressed with WT ELAVL1 or mutated ELAVL1) were measured by qPCR (n = 3). One-way ANOVA was performed: NS, ****P < 0.0001 (N); NS (O). Data represent the mean value ± SD from n biological replicates, or a representative immunoblot from n independent experiments. Source data are available for this figure: SourceData F7.

ELAVL1 is required for TNF/IFNγ-induced cell death. (A and C) HeLa (A) or HCT-116 (C) cells (parental, ELAVL1 KO clones #1 and #2 with different gRNA) were treated with TNF/IFNγ for 48 h (n = 3). Cell viability was determined by ATP levels. One-way ANOVA was performed: ****P < 0.0001. (B and D) The protein expression of ELAVL1 in indicated cells was analyzed by immunoblotting (n = 2). (E and G) HeLa (E) or HCT-116 (G) cells (parental, ELAVL1 KO, ELAVL1 KO stably expressed with WT ELAVL1 or mutated ELAVL1) were treated with TNF/IFNγ for 48 h (n = 3). Cell viability was determined by ATP levels. One-way ANOVA was performed: NS, ****P < 0.0001. (F and H) The protein expression of ELAVL1 in indicated cells was analyzed by immunoblotting (n = 2). (I) The binding interface between ELAVL1 and RNA in the ELAVL1-RNA complex (PDB 4ED5) is shown. Amino acids critical for the RNA binding of ELAVL1 are highlighted. (J) HeLa cells (parental, ELAVL1 KO clones #1 and #2 with different gRNA) were treated with IFNγ for 24 h (n = 3). The mRNA levels of CASP8 were measured by qPCR. One-way ANOVA was performed: ***P = 0.0003, ****P < 0.0001. (K) HeLa cells (parental, CASP8 KO, ELAVL1 KO clones #1 and #2 with different gRNA) were treated with IFNγ for 24 h. The expression of indicated proteins was analyzed by immunoblotting (n = 4). (L and M) HeLa cells (parental and ELAVL1 KO) were treated with actinomycin D for the indicated time. The mRNA levels of CASP8 (L) and CYLD (M) were measured by qPCR (n = 3). The mRNA levels in HeLa cells before the addition of actinomycin D (0 h) were set as 100%. Two-tailed t test was performed: **P = 0.0024 (L); NS (M). (N and O) The mRNA levels of CASP8 (N) and CYLD (O) in HeLa cells (parental, ELAVL1 KO, ELAVL1 KO stably expressed with WT ELAVL1 or mutated ELAVL1) were measured by qPCR (n = 3). One-way ANOVA was performed: NS, ****P < 0.0001 (N); NS (O). Data represent the mean value ± SD from n biological replicates, or a representative immunoblot from n independent experiments. Source data are available for this figure: SourceData F7.

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