ZFP36L1-deficient ASCs contain higher levels of GRK2.(A) GO terms enriched in gene set enrichment analysis performed on the genes differentially expressed (DE) in ZFP36L1-deficient and ZFP36L1-sufficient B cells activated with LPS. For each term, the numbers in parentheses are the numbers of all genes in the term followed by the adjusted P (padj) value. The numbers outside the gray bars represent the number of DE genes (padj < 0.05; fold change >1.5), and the numbers inside the blue bars represent the numbers of direct targets of ZFP36L1 identified by iCLIP (padj < 0.05), all overlapping with the corresponding GO terms. Under the conditions used, activated B cells undergo at most three cell divisions and do not contain any CD138highB220low cells. (B and C) Abundance of mRNA for S1pr1 gene for S1PR1 (B) and Adrbk1 gene encoding GRK2 kinase (C) expressed as FPKM in the LPS-activated B cells from Zfp36l1fl/fl CD79a+/+ (closed circles) and Zfp36l1fl/fl CD79aCre/+ (open squares) mice. Each symbol represents data from B cells of an individual mouse, lines represent means, and P values are adjusted by the Benjamini-Hochburg method calculated with DESeq2. (D) Conservation of the sequence surrounding cross-linked nucleotides in the 3′ UTR of Adrbk1 transcripts among primates and rodents. Cross-linked nucleotides were visualized using the UCSC Genome Browser (http://genome.ucsc.edu). In the gene diagram, the blue boxes denote exons, the thinner line denotes 3′ UTR, and the thin lines show introns. Arrows identify nucleotides cross-linked to ZFP36L1, and X indicates a nonconserved nucleotide. (E) An overlay of representative flow cytometry plots gated on CD138+NIP+IgG1+ cells from the spleens of Zfp36l1fl/fl CD79a+/+ and Zfp36l1fl/fl CD79aCre/+ mice analyzed 3 d after reimmunization. Rabbit polyclonal antibody against GRK2 was conjugated to Alexa Fluor 647. (F) A plot showing geometric mean fluorescent intensities (gMFIs) for the GRK2 signal in the population of splenic CD138+NIP+IgG1+ cells as in panel E for Zfp36l1fl/fl CD79a+/+ (closed circles) and Zfp36l1fl/fl CD79aCre/+ (open squares) mice. Each symbol represents data from an individual mouse, and lines represent means. Data for independent experiments are shown in Fig. S4 B (n ≥ 4 per experiment; the P value is for a Student’s t test).
Figure 4.

ZFP36L1-deficient ASCs contain higher levels of GRK2. (A) GO terms enriched in gene set enrichment analysis performed on the genes differentially expressed (DE) in ZFP36L1-deficient and ZFP36L1-sufficient B cells activated with LPS. For each term, the numbers in parentheses are the numbers of all genes in the term followed by the adjusted P (padj) value. The numbers outside the gray bars represent the number of DE genes (padj < 0.05; fold change >1.5), and the numbers inside the blue bars represent the numbers of direct targets of ZFP36L1 identified by iCLIP (padj < 0.05), all overlapping with the corresponding GO terms. Under the conditions used, activated B cells undergo at most three cell divisions and do not contain any CD138highB220low cells. (B and C) Abundance of mRNA for S1pr1 gene for S1PR1 (B) and Adrbk1 gene encoding GRK2 kinase (C) expressed as FPKM in the LPS-activated B cells from Zfp36l1fl/fl CD79a+/+ (closed circles) and Zfp36l1fl/fl CD79aCre/+ (open squares) mice. Each symbol represents data from B cells of an individual mouse, lines represent means, and P values are adjusted by the Benjamini-Hochburg method calculated with DESeq2. (D) Conservation of the sequence surrounding cross-linked nucleotides in the 3′ UTR of Adrbk1 transcripts among primates and rodents. Cross-linked nucleotides were visualized using the UCSC Genome Browser (http://genome.ucsc.edu). In the gene diagram, the blue boxes denote exons, the thinner line denotes 3′ UTR, and the thin lines show introns. Arrows identify nucleotides cross-linked to ZFP36L1, and X indicates a nonconserved nucleotide. (E) An overlay of representative flow cytometry plots gated on CD138+NIP+IgG1+ cells from the spleens of Zfp36l1fl/fl CD79a+/+ and Zfp36l1fl/fl CD79aCre/+ mice analyzed 3 d after reimmunization. Rabbit polyclonal antibody against GRK2 was conjugated to Alexa Fluor 647. (F) A plot showing geometric mean fluorescent intensities (gMFIs) for the GRK2 signal in the population of splenic CD138+NIP+IgG1+ cells as in panel E for Zfp36l1fl/fl CD79a+/+ (closed circles) and Zfp36l1fl/fl CD79aCre/+ (open squares) mice. Each symbol represents data from an individual mouse, and lines represent means. Data for independent experiments are shown in Fig. S4 B (n ≥ 4 per experiment; the P value is for a Student’s t test).

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