Supplemental Fig.1. The stable over-expression of FGFR4 in gastric cancer cells
were developed by transfection of LV5 containing human FGFR4 gene sequence.
(A) The construction of LV5 shuttle plasmid. (B) The development procedure of viral
particles containing LV5-NM_022963.2. (C-F) The transfection charts through
converted fluorescence microscope when the viral titers were 10-1 (1C), 10-2 (1D),
10-3 (1E) and 10-4 (1F), respectively.
Supplemental Fig.2. The different expression of FGFR4 could influence the
biological features of gastric cancer cells. Fig.2A-2C showed the invasion chamber of
MGC803 with Mock, NC and FGFR4-LV5, respectively. Fig.2D illustrated the
differences on invasion ability of MGC803 with Mock, NC and FGFR4-LV5
(Student’s t-test, *P < 0.05). Fig.2E revealed the differences concerning apoptosis rate
of MGC803 with Mock, NC and FGFR4-LV5 (Student’s t-test, *P < 0.05). Fig.2F
showed the expressions of different related molecules by western blot in BGC823
cells with Mock, NC and FGFR4-LV5.
Supplemental Fig.3.
The viability and proliferation ability of MGC803 cells with
different expression of FGFR4 were illustrated when treated with the single and
combination of 5-Fu and PD. Fig.3A-3C showed the viability of MGC803 treated by
5-Fu (3A), PD (3B) and 5-Fu plus PD (3C). Fig.3D and 3E displayed the proliferation
of MGC803 with NC (3D) and FGFR4-LV5 (3E) when treated by control, 5-Fu, PD
and 5-Fu plus PD (One-way ANOVA and Dunnett’s test, *P < 0.05). Figure 3F-3I
revealed the proliferation of MGC803 with NC and FGFR4-LV5 when treated by
control (3F), 5-Fu (3G), PD (3H) and PD plus 5-Fu (3I), which the statistical
differences were significant (Student’s t-test, *P<0.05). All experiments were
performed in triplicate and repeated at least three times.
Supplemental Fig.4. The apoptosis rates of MGC803 cells with different expression
of FGFR4 were shown when treated by the single and combination of 5-Fu and PD.
Fig. 4A-4D illustrated the apoptosis charts of MGC803 treated with control (4A),
5-Fu (4B), PD (4C) and 5-Fu plus PD (4D), which the statistical differences were
significant, especially when treated by 5-Fu plus PD (Fig.4E, *P < 0.05, **P < 0.01).
Compared with Mock and NC, the apoptosis rates of MGC803 with FGFR4-LV5
were much lower when treated with 5-Fu and 5-Fu plus PD while no difference in PD
group (Fig.4F, *P < 0.05, **P < 0.01). All experiments were performed in triplicate
and repeated at least three times.
Supplemental Fig.5. The expressions of related molecules including signal pathway
(erk and p-erk), apoptosis (Bcl-xl, Caspase-3 and Cleaved Caspase-3) and
proliferation (PCNA) by western blot were revealed in MGC803 cells with different
expression of FGFR4 when treated by the single and combination of 5-Fu and PD.
Fig.5A-5C displayed the expressions of related molecules in MGC803 with Mock,
NC and FGFR4-LV5 when treated by 5-Fu (Fig.5A), PD (Fig.5B) and PD plus 5-Fu
(Fig.5C), respectively. Fig.5D illustrated the expressions of related molecules in
MGC803 when treated by control, 5-Fu, PD and PD plus 5-Fu.