Figure 4

Transcriptional regulation of MMP12 by hnRNP K. (A) Schematic diagrams of the utilized reporter constructs, which contained 5' serial deletions of the MMP12 promoter. (B) NPC-TW02 cells were pretreated with control siRNA (C) or hnRNP K-targeting siRNA (K) for 24 h and then transfected with pGL3-basic (pGL3) with or without 5' serial deletions of the promoter sequence of the MMP 12. Firefly and Renilla luciferase activities were determined at 24 h post-transfection. *P < 0.05. (C) DNA pull-down assays were performed using nuclear extracts isolated from NPC-TW02 cells and 5' biotin-labeled probes corresponding to the −42/+97 (−42 to +97) or +2/+97 (+2 to +97) regions of the MMP12 promoter. The hnRNP K levels in the immunoprecipitates and 1% inputs were determined by Western blotting. (D) Chromatin immunoprecipitation was carried out using nuclear extracts from NPC-TW02 cells and an antibody against hnRNP K, followed by quantitative PCR of a sequence within the MMP12 promoter region (−95 to −20). Mouse IgG immunoprecipitation was done as a negative control. *P < 0.01.