Pharmacological Data Analysis Report of MTT and Plaque Assay

This report presents a comprehensive analysis of pharmacological data, specifically focusing on MTT and plaque assays. The report includes an overview of the methods used, including the use of Vero E6 cells, and the data obtained from the assays. The data is presented in the form of tables and graphical representations, which provides a detailed understanding of the results. The report covers the cellular analysis, the study of the expression of the MDR gene, and the kinetics of the 3-O-methylglucose uptake and efflux. The report also examines the plaque-forming units with time, the comparison of the normalized and non-normalized replicates, and the impact of the passage number. The report also covers the results of the study of the expression of the MDR gene. The report provides insights into the application of these assays in drug discovery and research.

Running head: PHARMACOLOGY
DATA ANALYSIS OF MTT AND PLAQUE ASSAY
Name of the Student
Name of the University
Author Note

1PHARMACOLOGY
MTT assay analysis
MTT assay is defined as a colorimetric assay which is used for accessing the
metabolic activity of cell. MTT is a yellow tetrazole which is reduced to purple formazan
inside living cells. After dissolution of the color, the absorbance is read in a
spectrophotometer.
Table 1 (Fold amplification of mdr (multi drug resistant) gene detected by RT PCR)
Normalized mdr
fold
Unnormalized mdr
fold
RNA fold
dilution
14 8 1
16 9 2
Graphical analysis
Normalized mdr fold Unnormalized mdr fold RNA fold dilution
0
2
4
6
8
10
12
14
16
18
Normalized vs Unnormalized fold
Series1 Series2
Fig 1: The mRNA which was 1 or 10 fold diluted for RT-PCR for mdr1 was compared
without normalization or by the normalization of beta 2 microtubule. Error bars have not
been considered for this analysis.

2PHARMACOLOGY
Table 2 (Cellular metabolic rates- normalized MTT assay)
Metabolic rate
(MCE)
Absorbance at 540
nm
10 0.15
20 0.21
30 0.3
40 0.4
Graphical analysis
10 20 30 40
0
0.05
0.1
0.15
0.2
0.25
0.3
0.35
0.4
0.45
Absorbance at 540 nm
Absorbance at 540 nm
Fig 2: Absorbance on Y axis increasing with cellular metabolic rate on X axis (Normalized
MTT assay data)

3PHARMACOLOGY
10 20 30 40
0
0.05
0.1
0.15
0.2
0.25
0.3
0.35
0.4
0.45
Normalized metabolic rates
Absorbance at 540 nm
Fig 3: Bar graph analysis showing the same as shown in figure 2.
Table 3 (Kinetics of 3OMG uptake and efflux on hepatocytes)
%Control Normalized Unnormalized
Glucose 100 101
Sucrose 98 80
D glucose 70 80
2DG 90 70
30MG 70 85

4PHARMACOLOGY
Graphical analysis
Glucose Sucrose D glucose 2DG 30MG
0
20
40
60
80
100
120
Normalized vs Unnormalized uptake levels
Normalized Unnormalized
Fig 4: Kinetics of 3OMG uptake and efflux on hepatocytes (bar graph analysis)
Glucose Sucrose D glucose 2DG 30MG
0
20
40
60
80
100
120
Normalized Unnormalized
Fig 5: Line graph for the kinetics of 3OMG uptake and efflux on hepatocytes

5PHARMACOLOGY
MTT assay 12 plates
MTT assay
plates
Absorbance
1 0.09
2 0.11
3 0.21
4 0.19
5 0.1
6 0.08
7 0.31
8 0.4
9 0.41
10 0.51
11 0.6
12 0.7
Graphical analysis
1 2 3 4 5 6 7 8 9 10 11 12
0
0.2
0.4
0.6
0.8
Absorbances
Absorbances
Line graph of the previous table

6PHARMACOLOGY
Plaque assay
Fig: 6: 9 well plate plaque assay
Table 4 (Plaque forming unit vs time)
Time (hours) Passage 29 Passage 34
24 80000 90000
48 60000 70000
72 58000 70000
Vero cells plated 48
hours before assay
Vero E6 cells plated
before assay (clear
plaques)
E6 cells plated 72
hours before assay

7PHARMACOLOGY
24 48 72
0
10000
20000
30000
40000
50000
60000
70000
80000
90000
100000
Comparison graph (normalized data)
Passage 29 Passage 34
Fig 7: EBOV titers were similar to ATCC passage cells. Each bar represents five replicates.
Fig 8: Filo-virus plaques produced on Vero E6 cells from two sources are similar in
Appearance and titer

8PHARMACOLOGY
Table 5 (Normalized and non normalized replicates vs PFU)
Replicates PFU/mL
Normalised 89000
NonNormalised 90000
Normalized 85000
NonNormalized 80000
Graphical analysis
Normalised NonNormalised Normalized NonNormalized
74000
76000
78000
80000
82000
84000
86000
88000
90000
92000
PFU/mL
Figure 9: Normalized and non normalized replicates vs PFU

9PHARMACOLOGY
Normalised NonNormalised Normalized NonNormalized
0
10000
20000
30000
40000
50000
60000
70000
80000
90000
100000
Line graph PFU/mL
Figure 10: Normalized and non normalized replicates vs PFU