Assistance of PLAG1 nucleus shuttling by KPNA2. (a) The association of KPNA2 and PLAG1 was assayed by Co-IP, protein samples not proceeding to IP was designated as input and samples pulled down by IgG antibody was used as negative control. Unassociated protein ACTB was examined to exclude unspecific bind by KPNA2 antibody. (b) The expression of KPNA2 (left panel) and PLAG1 (right panel) total protein in control Huh7 cells (GFP) or Huh7 cells transfected with KPNA2 expression plasmids (Clone1 and Clone2). (c) The expression of KPNA2 (left panel) and PLAG1(right panel) total protein in control SMMC7721 cells (Scramble) or SMMC7721 cells transfected with KPNA2 siRNAs (Si144 and Si467). (d) Nucleus accumulation of KPNA2 could be manipulated by KPNA2 expression plasmids and siRNAs. (e) The nucleus accumulation (up panel) and cytoplasm expression (down panel) of PLAG1 in SMMC7721 and Huh7 cells. ACTB and Lamin B antibody were applied for endogenous antibody for total and nuclearnucleus protein determination respectively. (f) In situ observation of the nucleus accumulation of PLAG1 in Huh7 cell line was investigated by immunocytochemistry. Nucleus was stained by DAPI. Cells with KPNA2 overexpression was marked by the white arrows. (g-h) Expression of transcriptional targets of PLAG1 in SMMC7721 and Huh7 cells. Data represents as mean ± s.d. ★ represents statistical significance.