Research Paper Volume 12, Issue 7 pp 5675—5692

STAT5A induced LINC01198 promotes proliferation of glioma cells through stabilizing DGCR8

Figure 4. LINC01198 interacted with DGCR8 stabilizing DGCR8. (A) Immunoblotting detection of DGCR8 in the LINC01198 pull-down complex. LINC01198 fragment (hereafter referred to as LINC01198) was labeled with the biotin; Non-sense RNA was labeled with the biotin whose length was similar to that of LINC01198 fragment; NC was the LINC01198 without biotin label. The molecular weight of DGCR8 approximates 120 kilodalton (kDa), suggested by manufacturer’s instruction that accompanies. (B) The PCR product of RNA immunoprecipitation (RIP), run by agarose gel electrophoresis with 12% separating gel. (C) qRT-PCR verified that LINC01198 was accumulated in DGCR8-precipatated protein sample. Two-tailed, independent sample T-test was used to analyze the significant difference (t=7.794, df=8, P<0.0001). (D) immunoblotting detection of DGCR8 stability in the presence of Cycloheximide (CHX) at 12.5μg/mL in T87G cells transfected with or without sh-LINC01198. gray density was quantitated by Image J (NIH, Bethesda, MA,USA). (E) Knock down efficiency of siRNA to DGCR8 at two different interfering sites, as evaluated by western-blot. Shown were the representative ones singled out from candidates out of three independent experiments. (F) Cluster analysis of heat map of miRNA microarray (SurePrint, homo, Angilent, USA) performed on T87G and Hs 684 glioma cells transfected with siRNA to DGCR8. Transfection with siRNA-scramble was set as control.