Figure 3. Role of menin in the down-regulation of INK4c.

A. Diagram of the human INK4c locus with exons depicted as boxes and coding domains shaded. The location of the transcriptional start sites (arrows) and primer sets P1-P6 used for ChIP analyses are indicated. B. ChIP of menin and H3K4me3 at the INK4c locus before and after treatment of IMR90 ER:RAS cells with OHT. Enrichment for each primer set was assessed by qPCR relative to a non-specific IgG and expressed as percentage of input. C and D. Comparison of MEN1 and INK4c RNA levels in proliferating (P) and senescent (S) populations of Hs68 (C) and Leiden (D) fibroblasts (same samples as in Fig. 2A and B). E. Relative levels of MEN1 and INK4c RNA in cells transduced with retroviral vectors encoding HA-tagged versions of p16^INK4a, p18^INK4c or p21^CIP1, as in Fig. 2C. F. Relative levels of INK4c, MEN1 and INK4a RNAs in Hs68 cells infected with a retrovirus encoding H-RAS^G12V or empty vector control (Vec), as in Fig. 2D. G. Relative levels of INK4c, MEN1 and INK4a RNAs following addition of tamoxifen (OHT) to BF cells expressing ER:H-RAS^G12V (as in Fig. 2E).