Fig. 6
From: Fstl1/DIP2A/MGMT signaling pathway plays important roles in temozolomide resistance in glioblastoma
Fstl1 overexpression inhibits and knockdown enhances DIP2A nuclear translocation. a Immunofluorescence assay was performed on D54 cells using anti-Fstl1 antibody (red) and anti-DIP2A antibody (green). Merge 1: Green + Red; Merge 2: Green + Red + Blue. Scale bar: 20 μm. b Co-IP was performed on total, membrane, cytoplasmic, and nuclear extracts prepared from D54 cells transfected with flag-Fstl1 using anti-flag antibody. c Immunofluorescence assay was performed on D54 cells transfected with Fstl1 or vector control, using anti-DIP2A antibody (green). Merge: Green + Blue. Scale bar: 20 μm. d The protein levels of DIP2A in the cytoplasm, nucleus, and membrane of D54 cells transfected with Fstl1 or vector control were detected by WB. Na + –K + –ATPase, GAPDH, and histone H3 were used as loading control for membrane, cytoplasm, and nucleus protein, respectively. e, f Co-IP was performed using ME and CE prepared from D54 cells transfected with Fstl1 or vector control using the indicated antibodies. g The protein levels of DIP2A in the cytoplasm, nucleus, and membrane of 05MG cells stably expressing shFstl1–7 or shCtrl were detected by WB. h, i Co-IP was performed using ME and CE prepared from GBM cells transfected with shFstl1–7 or shCtrl using the indicated antibodies. ME membrane extracts, CE cytoplasmic extracts, NE nuclear extracts. Student’s t tests were performed. Data are presented as mean ± SEM in three biological replicates (*P < 0.05, **P < 0.01)
