Extended Data Figure 9: PROX1 overexpression increases p300 binding and H3K9 acetylation at the CPT1A gene.

a, p300 enrichment around the human CPT1A gene in VECs (blue) and pLECs (green), as determined by ChIP-seq analysis of VECs transduced with a lentiviral vector expressing a control vector (GFP) or PROX1, respectively. Reads are binned as 100 bp intervals, and expressed per million reads (y axis). Displayed are averages of 2 independent ChIP-seq experiments. Top line: scheme of the CPT1A gene; grey blocks denote CPT1A exons and black blocks denote exons of adjacent genes. Bottom line: chromosomal location (human genome build GRCh37/hg19). The red arrowheads (A, B) above the chromosome location refer to statistically significant regions of PROX1 enrichment from Fig. 3g. b, c, p300 ChIP and qPCR for the PROX1-binding site at CPT1A ‘A’ (b) or CPT1A ‘B’ (c) in pLECs without (ctrl) and with CPT1A silencing (CPT1A^KD) versus VECs, values expressed as a percentage of input and relative to VECs (n = 3). d, H3K9ac enrichment around the human CPT1A gene in VECs (blue) and pLECs (green), as determined by ChIP-seq analysis of VECs transduced with a lentiviral vector expressing a control vector (GFP) or PROX1, respectively. Top line, scheme of the CPT1A gene; grey blocks denote CPT1A exons and black blocks denote exons of adjacent genes. Bottom line: chromosomal location (genome build hg19). The red arrowheads (A, B) above the chromosome location refer to statistically significant regions of PROX1 enrichment from Fig. 3g. q value is the FDR corrected P value. e, Quantitative PCR of the PROX1-binding regions ‘A’ and ‘B’ of the CPT1A gene (illustrated in d) on H3K9ac-ChIP samples from pLECs versus VECs, calculated as relative to the input and expressed relative to the values obtained for VECs (n = 3). f, g, Quantitation of the densitometry from PROX1-ChIP and PCR for the PROX1-binding site at CPT1A ‘A’ (f) or CPT1A ‘B’ (g) (identified in Fig. 3g) in pLECs without (ctrl) and with CPT1A silencing (CPT1A^KD) versus VECs, expressed as relative to input (n = 3). h, Quantitation of the densitometry from PROX1-ChIP and PCR for the PROX1-binding site at VEGFR3 in pLECs without (ctrl) and with CPT1A silencing (CPT1A^KD) versus VECs, expressed as relative to input (n = 3). i, p300 ChIP and qPCR for the PROX1-binding site at VEGFR3 in pLECs without (ctrl) and with CPT1A silencing (CPT1A^KD) versus VECs, values expressed as a percentage of input and relative to VECs (n = 3). Mean ± s.e.m. of n independent experiments. Statistical test: ANOVA and Bonferroni post-hoc test were used in multiple group comparisons. t-test was used for comparison of two groups. *P < 0.05; NS, not statistically significant.