A, Venn diagrams showing the overlap between the H3K27ac
peaks in B220+ cells from VavP-Bcl2/GFP tumors (n=4) or from
VavP-Bcl2/shCrebbp tumors (n=6) (left panel), or the
overlap between the H3K27ac peaks in MD901 cells transduced with either control
scramble shRNA (n=3) or shRNAs against CREBBP (n=6) (right
panel). B, Stacked bar plot representing the genomic distribution
of common and lost H3K27ac peaks between VavP-Bcl2/GFP and
VavP-Bcl2/shCrebbp tumor cells (left panel), or between
MD901 cells transduced with either control scramble or CREBBP
shRNAs (n=6). C, Normalized average H3K27ac read density plot at
loci identified as H3K27ac peaks in MACS-purified B220+ B cells from
VavP-Bcl2/GFP tumors. The black line represents the average
values in VavP-Bcl2/EV tumors (n=4), and the dark red line
represents the values in VavP-Bcl2/shCrebbp
tumors (n=6). Average values of peaks that exhibited more than 25%,
35%, 40%, and 50% reads loss in
VavP-Bcl2/shCrebbp tumors as compared to
VavP-Bcl2/GFP tumors are shown as lines in different shades
of red. * representing statistical significant loss of normalized H3K27ac read
density as determined by Kolmogorov-Smirnov test. D, Bar plot
representing ratio of the proportions of enhancer peaks or promoter peaks that
exhibited more than 25%, 35%, 40%, and 50% reads
loss in VavP-Bcl2/shCrebbp tumors as compared
to VavP-Bcl2/GFP tumors. E, Normalized average
H3K27ac read density plot at loci identified as H3K27ac peaks in scramble shRNA
transduced MD901 cells. The black line represents the average values in scramble
shRNA transduced MD901 cells (n=3), and the dark red line represents the values
in CREBBP shRNAs transduced MD901 cells (n=6). Average values
of peaks that exhibited more than 25%, 40%, 50%, and
70% reads loss in CREBBP KD cells as compared to
control scramble cells are shown as lines in different shades of red. *
representing statistical significant loss of normalized H3K27ac read density as
determined by Kolmogorov-Smirnov test. F, Bar plot representing
ratio of the proportions of enhancer peaks or promoter peaks that exhibited more
than 25%, 40%, 50%, and 70% reads loss in
CREBBP KD cells as compared to control scramble MD901
cells. G, UCSC read-density tracks of normalized H3K27ac ChIP-seq
reads at murine Cd86 locus in two representative
VavP-Bcl2/GFP tumors and two representative
VavP-Bcl2/shCrebbp tumors. Shaded areas
highlight the regions showed loss of H3K27ac in
VavP-Bcl2/shCrebbp tumors. H,
UCSC read-density tracks of normalized H3K27ac ChIP-seq reads at human
CD86 locus in two representative biological replicates of
control scramble (Scr) MD901 cells and two representative biological replicates
of CREBBP KD (shCREBBP) MD901 cells. Shaded
areas highlight the regions showed loss of H3K27ac in KD cells. I,
Pathways analysis of genes (n=1147) with > 25% reduction of
H3K27ac reads at enhancers in murine
VavP-Bcl2/shCrebbp tumors, or genes
(n=2928) with > 25% reduction of H3K27ac reads at enhancers in
CREBBP knockdown cells. Heatmap represents the BH-adjusted
p value of each geneset tested. J, UCSC read-density tracks of
normalized BCL6 (purple) and SMRT (orange) ChIP-seq reads in human tonsilar
GCBs, H3K27ac ChIP-seq reads in human tonsilar NBCs (red) and GCBs (blue), and
H3K27ac ChIP-seq in control scramble (Scr, green) and CREBBP KD
(shCREBBP, turquoise) MD901 cells at the human MHC II loci.
BCL6 and SMRT peaks determined by MACS2 are indicated by grey bars under the
read density track. Shaded areas highlight the enhancers that were bound by BCL6
and SMRT, and showed loss of H3K27ac in CREBBP KD cells.