hnRNP K levels are not influenced by hnRNP A1 expression. (A–C) RT-qPCR analysis of hnRNP A1 (A and B) and hnRNP K (C) mRNA levels in HEK293T cells overexpressing FLAG-hnRNP K (A) or with hnRNP K KD (B) or overexpressing hnRNP A1 (C). Data are normalized to ACTB. Statistical analyses were performed using one-way ANOVA followed by Tukey’s test (A and C) or Welch’s t test (B), respectively (n = 3 for each group). (D) WB analysis of HEK293T cells overexpressing FLAG-hnRNP A1. Quantification of hnRNP K is normalized to β-actin. Statistical analysis was performed using one-way ANOVA followed by Tukey’s test (n = 3 for each group). (E) Immunocytochemistry of HeLaS3 cells overexpressing FLAG-hnRNP K. FLAG (magenta), hnRNP A1 (green), and Hoechst (blue). Arrowheads and arrows indicate FLAG-positive and FLAG-negative cells, respectively. Scale bar: 10 μm. (F) WB analysis of HEK293T cells with hnRNP A1 KD. Quantification of hnRNP A1 and hnRNP K is normalized to β-actin. Statistical analyses were performed using Welch’s t test (n = 3 for each group). (G) RT-qPCR analysis of hnRNP K mRNA levels in HEK293T cells overexpressing FLAG-hnRNP A1. Data are normalized to ACTB. Statistical analysis was performed using Welch’s t test (n = 3 for each group). (H) IP of FLAG-hnRNP A1–overexpressing HEK293T cells. FLAG antibody was used for IP, and endogenous hnRNP K was detected by WB. An anti-IgG antibody was used as a negative control for IP. (I–K) Schematic diagram showing conditions with excess hnRNP K (I), hnRNP A1 (J), or hnRNP K coexpressed with hnRNP A1 (K). All graphs show the mean ± SEM. *P < 0.05, **P < 0.01. Source data are available for this figure: SourceData FS2.