(a,b) Relative proliferation of MDA-MB-468 control (with or without
sapienate) and FADS2 overexpression cells upon treatment 0.5 nM Merck Frosst Cpd
3j normalized to control (a: n=9; b: control n=10,
overexpression n=12). Two-way ANOVA with Tukey’s multiple
comparisons.
(c,d) Relative proliferation of HUH7 and A549 cells
(with or without sapienate) upon FADS2 knockdown with(out) 2 nM
Merck Frosst Cpd 3j normalized to control (c: control n=9;
shFADS2-1 n=6; shFADS2-2 n=6; d: n=6). Two-way ANOVA with Tukey
multiple comparisons (within different cell lines); one-way ANOVA with
Dunnett’s multiple comparisons (across different cell lines). Only
pair-wise comparisons are depicted.
(e,f) Representative images of hematoxylin and eosin
stain and relative area of resected tumor nodules derived from HUH7 control
(non-targeting shRNA) or FADS2 knockdown (shFADS2-2) orthotopic
liver xenografts in mice treated with vehicle or Merck Frosst Cpd 3j (1.5 mg per
kg twice daily per oral; p.o.; control+vehicle n=13; control+SCD inhibition
n=12; shFADS2-2+vehicle n=12, shFADS2-2+SCD inhibition n=14 of one experiment).
Masks in (e) show the tissue contour and indicating tumor (black), necrotic
(grey) and liver (white) area. Scale bar represents 1,000 µm in all
cases. Box blots in (f) show box extending from the 25th to
75th percentiles, whiskers indicating the minimum and maximum,
and a line indicating the mean. One-way ANOVA with Tukey’s multiple
comparisons.
Experiments were performed in low FBS (1%: HUH7; 0.5%: others) with treatment of
72 h. Error bars represent SD from mean of biological independent samples
(in vitro) or animals (in vivo), unless
stated otherwise.