Identification of Arid5a target transcripts in RTECs. (a) Experimental design. Inset: HK-2 cells were transfected with Flag-Arid5a and treated ± IL-17 for the indicated times. Flag-Arid5a was assessed in cell lysates by immunoblotting, representative of three independent experiments. (b) HK-2 cells were transfected with Flag-Arid5a, treated ± IL-17 for 3 h, and enrichment of the indicated mRNAs after RIP was assessed by qPCR. Each symbol represents an individual sample, pooled from three independent experiments. Results were normalized to untreated IgG RIP controls. Data are mean ± SEM, analyzed by one-way ANOVA with post-hoc Tukey’s test for multiple comparisons. (c) HK-2 cells were transfected with Flag-Arid5a, treated ± IL-17 for 3 h, and lysates were subjected to RIP-Seq with anti-Flag or IgG isotype controls. Heatmaps show selected inflammatory mRNAs from three independent replicates. (d) GSEA pathway prediction of RIP-Seq comparing untreated and IL-17–treated samples, FDR < 0.05, minimum gene IDs in category = 5. (e) HK-2 cells were transfected with Flag-Arid5a, treated ± IL-17 for 3 h, and lysates subjected to RIP. Enrichment of rRNAs corresponding to 40S (18S) and 60S ribosomes (5S, 5.8S, 28S) were determined by qPCR, normalized to untreated IgG RIP controls. Data show mean ± SEM, analyzed by one-way ANOVA with post-hoc Tukey’s test. Data were pooled from five independent experiments. *P < 0.05, ** < 0.01, *** < 0.001, **** < 0.0001. Source data are available for this figure: SourceData F5.
Figure 5.

Identification of Arid5a target transcripts in RTECs. (a) Experimental design. Inset: HK-2 cells were transfected with Flag-Arid5a and treated ± IL-17 for the indicated times. Flag-Arid5a was assessed in cell lysates by immunoblotting, representative of three independent experiments. (b) HK-2 cells were transfected with Flag-Arid5a, treated ± IL-17 for 3 h, and enrichment of the indicated mRNAs after RIP was assessed by qPCR. Each symbol represents an individual sample, pooled from three independent experiments. Results were normalized to untreated IgG RIP controls. Data are mean ± SEM, analyzed by one-way ANOVA with post-hoc Tukey’s test for multiple comparisons. (c) HK-2 cells were transfected with Flag-Arid5a, treated ± IL-17 for 3 h, and lysates were subjected to RIP-Seq with anti-Flag or IgG isotype controls. Heatmaps show selected inflammatory mRNAs from three independent replicates. (d) GSEA pathway prediction of RIP-Seq comparing untreated and IL-17–treated samples, FDR < 0.05, minimum gene IDs in category = 5. (e) HK-2 cells were transfected with Flag-Arid5a, treated ± IL-17 for 3 h, and lysates subjected to RIP. Enrichment of rRNAs corresponding to 40S (18S) and 60S ribosomes (5S, 5.8S, 28S) were determined by qPCR, normalized to untreated IgG RIP controls. Data show mean ± SEM, analyzed by one-way ANOVA with post-hoc Tukey’s test. Data were pooled from five independent experiments. *P < 0.05, ** < 0.01, *** < 0.001, **** < 0.0001. Source data are available for this figure: SourceData F5.

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