Treatment of A375 cells with PA (15 μM) for different time points iTRAQ Labeling Pool, SCX,...
5
Treatment of A375 cells with PA (15 μM) for different time points iTRAQ Labeling Pool, SCX, Desalting and Offline 2DLC MS/MS analysis Cell Lysis, Reduction, Alkylation, and Tryptic Digestion 0 h 4 h 12h 24h 0 h 4 h 12h 24h 113 114 115 116 117 118 119 121 Biological replicate 1 Biological replicate 2 Data Analysis Supporting Information Figure S1. (A) Schematic representation of the experimental design of the iTRAQ-based quantitative proteomics study. (B) Schematic representation of the data analysis pipeline of the proteomics results. A B porting Information Figure S1
-
Upload
bruce-short -
Category
Documents
-
view
225 -
download
0
description
Supporting Information Figure S3. (A) Enlargement of mitochondria of PA-treated SK-MEL-28 melanoma cells. Arrows showing vesicles accumulated upon PA treatment. Images were acquired using Cellomics ArrayScan HCS Reader. (B) Decrease abundance of COX IV and loss of MMP in PA-treated SK-MEL-28 melanoma cells. Representative images of three independent experiments were shown. Average fluorescent intensities of COX IV and MMP dye were quantified using CHP algorithm on the HCS platform. (C) Upregulation of HSPA5 and CHOP protein in PA-treated SK-MEL-28 melanoma cells. Data are expressed as means ± SEM of three independent experiments. Statistical significance is expressed as ***, P < 0.001; **, P < 0.01 versus untreated control. Supporting Information Figure S3
Transcript of Treatment of A375 cells with PA (15 μM) for different time points iTRAQ Labeling Pool, SCX,...
Treatment of A375 cells with PA (15 μM) for different time points
iTRAQ Labeling
Pool, SCX, Desalting and Offline 2DLC MS/MS analysis
Cell Lysis, Reduction, Alkylation, and Tryptic Digestion
0 h 4 h 12h 24h 0 h 4 h 12h 24h
113 114 115 116 117 118 119 121
Biological replicate 1 Biological replicate 2
Data Analysis
Supporting Information Figure S1. (A) Schematic representation of the experimental design of the iTRAQ-based quantitative proteomics study. (B)
Schematic representation of the data analysis pipeline of the proteomics results.
A
B
Supporting Information Figure S1
Supporting Information Figure S2 (A) Cell viabilities of A375 melanoma cells after 24 h treatment with PA as determined by MTT assay. (B) Accumulation of vesicles
on PA-treated A375 cells observed under inverted light microscope. (C) Real-time growth profile of A375 cells in response to treatment with PA.
60 µM 30 µM 15 µM 7.5 µM
3.75 µM 1.88 µM Untreated Control
A
-0.5 0.0 0.5 1.0 1.5 2.00
20
40
60
80
100
120
Log (M)
Cel
l Via
bilit
y (%
)B
CTRL PA (15 μM)
C
Supporting Information Figure S2
Supporting Information Figure S3. (A) Enlargement of mitochondria of PA-treated SK-MEL-28 melanoma cells. Arrows showing vesicles
accumulated upon PA treatment. Images were acquired using Cellomics ArrayScan HCS Reader. (B) Decrease abundance of COX IV and loss
of MMP in PA-treated SK-MEL-28 melanoma cells. Representative images of three independent experiments were shown. Average fluorescent
intensities of COX IV and MMP dye were quantified using CHP algorithm on the HCS platform. (C) Upregulation of HSPA5 and CHOP protein in
PA-treated SK-MEL-28 melanoma cells. Data are expressed as means ± SEM of three independent experiments. Statistical significance is
expressed as ***, P < 0.001; **, P < 0.01 versus untreated control.
Supporting Information Figure S3
Supporting Information Figure S4. (A) Correlation plot for the two biological replicates of 0 h control (N=4092). (B)
Correlation plot for the two biological replicates of 0 h control (N=296).
Supporting Information Figure S4
Supporting Information Figure S5. Correlation plots for the two biological replicates of (A) 4 h post-treatment with PA
(N=35). (B) 12 h post-treatment with PA (N=145). (C) 24 h post-treatment with PA (N=245).
Supporting Information Figure S5
