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. 2020 Apr 17;15(4):e0231450. doi: 10.1371/journal.pone.0231450

Fig 5. HNRNPU regulates P/I-induced IL-6 mRNA expression.

Fig 5

(A–C) HeLa cells were transfected with siRNAs, incubated for 72 hours, and then treated with P/I for 4 hours (B) or 24 hours (A, C) and harvested. HNRNPU protein expression was determined by Western blotting with actin as a loading control (A). IL-6 mRNA expression was examined using a TaqMan assay and normalized with RPLP0 (B). (C) IL-6 protein secreted in the condition medium was quantified by ELISA (R&D Systems). Statistically significant differences are marked with *** (p < 0.005, Welch’s t-test). (D) HeLa cell lysates that had been stimulated with P/I for 4 hours were subjected to a RIP assay using the anti-HNRNPU antibody or normal mouse IgG as a negative control. Co-immunoprecipitated mRNAs were reverse transcribed with Superscript III, and co-immunoprecipitated IL-6 and RPLP0 mRNA were quantified using a TaqMan assay.