Figure 6

miR-375 expression is regulated by N-myc. A. Representative western blot of N-myc protein - SH-SY5Y (1), SMS-LHN (2), BE(2)-M17V (3), LA1-55n (4), KCN-83n (5), SK-N-BE(1)n (6), CB-JMN (7), BE(2)-C (8), SK-N-LD (9), SK-N-HM (10), SH-EP1 (11), SMS-KCNs (12), and LA1-5s (13). B. Changes in miR-375 levels, compared to U6, in N-myc antisense-transfected (As) LA1-55n cells and N-myc sense-transfected SH-SY5Y cells (sense) compared to vector-transfected (vec) controls. C. Chromatin immunoprecipitation analysis of N-myc regulation of miR-375. DNA-protein complexes cross-linked with formaldehyde were isolated and sonicated (Input DNA). Aliquots (1% of input) were immunoprecipitated with antibodies to N-myc or RNA polymerase II (Pol II) or with goat anti-rabbit secondary antibody. DNA was amplified by PCR with primers specific to miR-375 E-box sequence (1), Hook1 (2), and GAPD (3). Note that primers specific for miR-375 and GAPD E-box sequences gave a band indicating the association of N-myc with those genes.