Figure legends
Figure 1. C26-derived CM induces atrophy in C2C12
myotubes
Schematic representation of the
processes involved in differentiation of C2C12 myoblast into myotubes
and the subsequent treatment with CM isolated from C26 colon cancer
cells. (B ) Representative images obtained from
immunofluorescence (40×) highlighting the myotube diameter upon
treatment with C26-derived CM. Scale bars represent 20 μm.
(C ) Quantification of myotube diameter (n=3). (D )
Western blot (80µg) showing a decrease in the expression of MyHC in
C2C12 treated with C26-derived CM. β-actin was used as a loading
control to confirm equal loading of protein. (E ) Metabolic
activity of untreated and C26-dericed CM treated C2C12 myotubes at 24,
48, and 72 h. Absorbance was measure at 490 and 630 nm (n = 3). All
data represented as mean ± s.e.m.; p-values as determined by
two-tailed unpaired students’ t-test.Figure 2. Protein profile of C2C12 upon treatment
with C26-derived CM
Schematic diagram depicting the step involved in mass
spectrometry-based proteomics. (B) Venn diagram showing the
total number of proteins identified between untreated and C26-dervied
CM treated C2C12. (C) Quantitative Venn diagram portraying
differential abundance of proteins identified in untreated and
C26-dervied CM treated C2C12 cell lysates. (D) Volcano plot
displaying the scatter of proteins identified in untreated and
C26-dervied CM treated C2C12 cells. Curved lines represent
significance threshold (n = 3). (E ) Heatmap showing
statistically significant proteins with fold change greater than ≥2
identified in untreated and C26-dervied CM treated C2C12 cells.
Figure 3. Enrichment analysis of C26-derived CM treated
C2C12
Gene enrichment analysis highlighting the biological processes that
are enriched in C26-dervied CM treated C2C12 myotubes. (B)Gene enrichment analysis highlighting the biological processes that
are depleted in C26-dervied CM treated C2C12 myotubes. (C )
Western blot (60 μg) showing the abundance of BiP and calnexin in
C2C12 upon C26 CM treatment. β-actin was used as a loading control to
confirm equal loading of protein. (D ) Relative intensity of
Bip and calnexin in C26-derived CM treated C2C12 compared to untreated
C2C12.n = 3; all data represented as mean ± s.e.m.; p-values as
determined by two-tailed unpaired students’ t-test.