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

From: The role of fatty acid β-oxidation in lymphangiogenesis

Extended Data Figure 9

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 (CPT1AKD) 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 (CPT1AKD) 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 (CPT1AKD) 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 (CPT1AKD) 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.