LIFE01200604005 Shri Somnath Ghosh - Homi Bhabha National ...

More documents

Recommendations

Info

CHAPTER 2 MATERIALS AND METHODS 2.11 Measurement of NO production in EL-4 cells Control unirradiated cells, cells exposed to radiation and the cells exposed to different conditioned media as mentioned above were cultured for 24 h at 37 0 C in 5%CO 2 / 95% air. The production of NO in the culture supernatants was measured by assaying NO - 2 using colorimetric Griess reaction [230]. In brief, 100 µl of culture supernatants was incubated with an equal volume of Griess reagent [1% sulfanilamide and 0.1% N-(1-naphthyl-ethylenediamine) in 2.5% phosphoric acid; Sigma, USA] at room temperature for 10 min in a 96 well plate. The absorbance at 550 nm was measured using a microplate reader (Fluostar Optima, Biotron Healthcare). Absorbance measurements were converted to µ moles of NO 2 - using a standard curve of NaNO 2. 2.12 Measurement of Apoptosis by DNA fragmentation Apoptosis in control unirradiated cells, cells exposed to radiation and the cells exposed to different conditioned media as mentioned above was measured by DNA fragmentation assay [231]. After transfer of medium from irradiated cells, all the above mentioned groups of EL-4 cells were cultured for 24 h at 37 0 C in 5%CO 2 , 95% air. The cells were harvested and lysed in lysis buffer (10 mM EDTA, 50 mM Tris–HCl, pH 8.0, 0.5% sodium lauryl sarcosine) containing the 100 mg/ml proteinase K at 55 o C for 2 h. The DNA was extracted with phenol/chloroform and precipitated with ethanol. The RNA was removed by RNAse A treatment and DNA was electrophorased on agarose gel to visualize the fragmented DNA. 91
CHAPTER 2 MATERIALS AND METHODS 2.13 Transfection with ShRNA MLH1, Rad52 and Control shRNA plasmids were procured from Santa cruz biotechnology (Cat No. sc-35943-SH, sc-37399-SH and sc-108060 respectively). In a six well tissue culture plate, A549 cells were grown to 50-70% confluency in antibiotic-free normal growth medium supplemented with FBS and transfection was carried out as per supplier’s instruction. 48 hours post-transfection, medium was aspirated and replaced with fresh medium containing puromycin (2µg/ml) for selection of stably transfected cells. Semi-quantitative RT-PCR was performed to monitor MLH1 and Rad52 gene expression knockdown using gene specific primers (Table 1). 2.14 Statistical Analysis The data were imported to excel work sheets, and graphs were made using Origin version 5.0. One-way ANOVA with Tukey–Kramer Multiple Comparisons as post-test for P < 0.05 used to study the significant level. Data were insignificant at P > 0.05. Each point represented as mean±S.E. (standard error of the mean). 92
Page 1:
RADIATION INDUCED SIGNALING IN MAMM
Page 4 and 5:
DECLARATION I, hereby declare that
Page 6 and 7:
CONTENTS Page No. SYNOPSIS………
Page 8 and 9:
2.11 Measurement of NO production
Page 10 and 11:
PREAMBLE SYNOPSIS Ionising radiatio
Page 12 and 13:
SYNOPSIS Aims and Objectives: To St
Page 14 and 15:
SYNOPSIS 2.6 Immunofluorescence sta
Page 16 and 17:
SYNOPSIS ATM gene expression, which
Page 18 and 19:
SYNOPSIS Percentage Survival 100 80
Page 20 and 21:
SYNOPSIS Ratio of Rad52 to Actin Ra
Page 22 and 23:
Relative Phosphorylation 45 40 35 3
Page 24 and 25:
SYNOPSIS Fig. 3.3A Clonogenic Cell
Page 26 and 27:
SYNOPSIS (A) Av No. of phospho BRCA
Page 28 and 29:
SYNOPSIS Percentage Survival 120 10
Page 30 and 31:
SYNOPSIS Fig.3.4B. Existance of cro
Page 32 and 33:
SYNOPSIS Fig. 3.4EDNA damage in EL-
Page 34 and 35:
SYNOPSIS subsequent cytotoxicity ca
Page 36 and 37:
SYNOPSIS [4] Hall, E. J. Radiobiolo
Page 38 and 39:
CHAPTER 1 INTRODUCTION INTRODUCTION
Page 40 and 41:
CHAPTER 1 INTRODUCTION 15.8% 9.56%
Page 42 and 43:
CHAPTER 1 INTRODUCTION far apart an
Page 44 and 45:
CHAPTER 1 INTRODUCTION and are more
Page 46 and 47:
CHAPTER 1 INTRODUCTION 1.3 DNA dama
Page 48 and 49:
CHAPTER 1 INTRODUCTION associates w
Page 50 and 51: CHAPTER 1 INTRODUCTION are unable t
Page 52 and 53: CHAPTER 1 INTRODUCTION Perhaps one
Page 54 and 55: CHAPTER 1 INTRODUCTION occurs at DS
Page 56 and 57: CHAPTER 1 INTRODUCTION related prot
Page 58 and 59: CHAPTER 1 INTRODUCTION damage must
Page 60 and 61: CHAPTER 1 INTRODUCTION At the prese
Page 62 and 63: CHAPTER 1 INTRODUCTION predominant
Page 64 and 65: CHAPTER 1 INTRODUCTION provide a me
Page 66 and 67: CHAPTER 1 INTRODUCTION hypersensiti
Page 68 and 69: CHAPTER 1 INTRODUCTION cycle-specif
Page 70 and 71: CHAPTER 1 INTRODUCTION 1.4 Overview
Page 72 and 73: CHAPTER 1 INTRODUCTION 1.4.1 Radiat
Page 74 and 75: CHAPTER 1 INTRODUCTION interaction
Page 76 and 77: CHAPTER 1 INTRODUCTION p90S6 kinase
Page 78 and 79: CHAPTER 1 INTRODUCTION CD4 (formerl
Page 80 and 81: CHAPTER 1 INTRODUCTION unrepaired d
Page 82 and 83: CHAPTER 1 INTRODUCTION well to the
Page 84 and 85: CHAPTER 1 INTRODUCTION 1.6 Radiatio
Page 86 and 87: CHAPTER 1 INTRODUCTION becomes pote
Page 88 and 89: CHAPTER 2 MATERIALS AND METHODS MAT
Page 90 and 91: CHAPTER 2 MATERIALS AND METHODS 2.2
Page 92 and 93: CHAPTER 2 MATERIALS AND METHODS res
Page 94 and 95: CHAPTER 2 MATERIALS AND METHODS Arr
Page 96 and 97: CHAPTER 2 MATERIALS AND METHODS PBS
Page 98 and 99: CHAPTER 2 MATERIALS AND METHODS the
Page 102 and 103: CHAPTER 3 RESULTS RESULTS 93
Page 104 and 105: CHAPTER 3 RESULTS fractionated regi
Page 106 and 107: CHAPTER 3 RESULTS On comparison of
Page 108 and 109: CHAPTER 3 RESULTS Table 3.1.2A List
Page 110 and 111: CHAPTER 3 RESULTS 80 NM_002185 IL7R
Page 112 and 113: CHAPTER 3 RESULTS SAA2 149 AU134977
Page 114 and 115: CHAPTER 3 RESULTS 221 LOC643167 RNA
Page 116 and 117: CHAPTER 3 RESULTS 299 BF244402 FBXO
Page 118 and 119: CHAPTER 3 RESULTS CDK4) 57 AU152107
Page 120 and 121: CHAPTER 3 RESULTS 134 AL045793 METT
Page 122 and 123: CHAPTER 3 RESULTS 58 AF237813 ABAT
Page 124 and 125: CHAPTER 3 RESULTS 127 AI809749 ORMD
Page 126 and 127: CHAPTER 3 RESULTS 31 BE615699 UNC84
Page 128 and 129: CHAPTER 3 RESULTS 47 AF026303 SULT1
Page 130 and 131: CHAPTER 3 RESULTS 117 BF062193 CYor
Page 132 and 133: CHAPTER 3 RESULTS 187 AL036662 ---
Page 134 and 135: CHAPTER 3 RESULTS 52 AV686514 EMP2
Page 136 and 137: CHAPTER 3 RESULTS exposed to 10 Gy
Page 138 and 139: CHAPTER 3 RESULTS Fig.3.1.4 Gene ex
Page 140 and 141: CHAPTER 3 RESULTS Fig. 3.1.6 Radiat
Page 142 and 143: CHAPTER 3 RESULTS Fig. 3.1.7 Transl
Page 144 and 145: CHAPTER 3 RESULTS 3.1.9 Stabilizati
Page 146 and 147: CHAPTER 3 RESULTS 23±1.5% (Fig. 3.
Page 148 and 149: CHAPTER 3 RESULTS whereas only the
Page 150 and 151:
CHAPTER 3 RESULTS Fig.3.2.2 Gene ex
Page 152 and 153:
CHAPTER 3 RESULTS Fig.3.2.4 Gene ex
Page 154 and 155:
CHAPTER 3 RESULTS with equal doses
Page 156 and 157:
CHAPTER 3 RESULTS Fig.3.3.1.2 Forma
Page 158 and 159:
CHAPTER 3 RESULTS Fig.3.3.1.3 Phosp
Page 160 and 161:
CHAPTER 3 RESULTS (7.5±0.64) (Fig.
Page 162 and 163:
Page 164 and 165:
CHAPTER 3 RESULTS Fig.3.3.1.7b Phos
Page 166 and 167:
CHAPTER 3 RESULTS 3.3.2 Oxygen beam
Page 168 and 169:
CHAPTER 3 RESULTS in all the cells.
Page 170 and 171:
CHAPTER 3 RESULTS 3.3.2.3 Radiation
Page 172 and 173:
Page 174 and 175:
Page 176 and 177:
CHAPTER 3 RESULTS Fig.3.3.2.7 Apopt
Page 178 and 179:
CHAPTER 3 RESULTS PMA stimulated U9
Page 180 and 181:
CHAPTER 3 RESULTS Fig.3.4.2. Exista
Page 182 and 183:
CHAPTER 3 RESULTS up-regulation of
Page 184 and 185:
CHAPTER 3 RESULTS Fig.3.4.3.2 NO pr
Page 186 and 187:
CHAPTER 3 RESULTS 3.4.3.4 Apoptotic
Page 188 and 189:
CHAPTER 3 RESULTS 3.4.4 Bystander e
Page 190 and 191:
CHAPTER 3 RESULTS Fig.3.4.4b DNA da
Page 192 and 193:
CHAPTER 3 RESULTS Fig.3.4.5a Gene e
Page 194 and 195:
CHAPTER 4 DISCUSSION DISCUSSION 185
Page 196 and 197:
CHAPTER 4 DISCUSSION directly expos
Page 198 and 199:
CHAPTER 4 DISCUSSION dose is delive
Page 200 and 201:
CHAPTER 4 DISCUSSION A clear cause
Page 202 and 203:
CHAPTER 4 DISCUSSION 4.2 Proton bea
Page 204 and 205:
CHAPTER 4 DISCUSSION Although the e
Page 206 and 207:
CHAPTER 4 DISCUSSION Similar number
Page 208 and 209:
CHAPTER 4 DISCUSSION the mechanism
Page 210 and 211:
CHAPTER 4 DISCUSSION Thus unlike th
Page 212 and 213:
CHAPTER 4 DISCUSSION different from
Page 214 and 215:
CHAPTER 4 DISCUSSION upregulates ma
Page 216 and 217:
CHAPTER 4 DISCUSSION Numerous studi
Page 218 and 219:
CHAPTER 4 DISCUSSION inhibition of
Page 220 and 221:
CHAPTER 4 DISCUSSION The survival o
Page 222 and 223:
CHAPTER 5 REFERENCES [1] Khan, F. T
Page 224 and 225:
CHAPTER 5 REFERENCES [19] Woo, R. A
Page 226 and 227:
CHAPTER 5 REFERENCES [35] Nghiem, P
Page 228 and 229:
CHAPTER 5 REFERENCES [52] McKay, B.
Page 230 and 231:
CHAPTER 5 REFERENCES [69] Cary, R.
Page 232 and 233:
CHAPTER 5 REFERENCES [84] Uziel, T.
Page 234 and 235:
CHAPTER 5 REFERENCES [98] Liu, Y.;
Page 236 and 237:
CHAPTER 5 REFERENCES [111] Fei, P.;
Page 238 and 239:
CHAPTER 5 REFERENCES [127] Frank, K
Page 240 and 241:
CHAPTER 5 REFERENCES [141] Pierce,
Page 242 and 243:
CHAPTER 5 REFERENCES [156] Roots, R
Page 244 and 245:
CHAPTER 5 REFERENCES [171] Xia, Z.;
Page 246 and 247:
CHAPTER 5 REFERENCES (MAP) kinase c
Page 248 and 249:
CHAPTER 5 REFERENCES phosphorylatio
Page 250 and 251:
CHAPTER 5 REFERENCES [211] Anderson
Page 252 and 253:
CHAPTER 5 REFERENCES [229] Konca, K
Page 254 and 255:
CHAPTER 5 REFERENCES [245] Kastan,
Page 256 and 257:
CHAPTER 5 REFERENCES [261] Miralbel
Page 258 and 259:
CHAPTER 5 REFERENCES [278] Weizman,
Page 260 and 261:
CHAPTER 5 REFERENCES [294] Zhou, H.
Page 262 and 263:
PUBLICATIONS AND PRESENTATIONS Publ
Page 264 and 265:
Mutation Research 729 (2012) 61-72
Page 266 and 267:
S. Ghosh, M. Krishna / Mutation Res
Page 268 and 269:
Page 270 and 271:
Page 272 and 273:
Page 274 and 275:
Page 276 and 277:
Page 278 and 279:
12 S. Ghosh et al. / Mutation Resea
Page 280 and 281:
Page 282 and 283:
Page 284 and 285:
Page 286 and 287:
Page 288 and 289:
192 S. Ghosh et al. / Mutation Rese
Page 290 and 291:
Page 292 and 293:
Page 294 and 295:
Page 296 and 297:
Cancer Invest Downloaded from infor
Page 298 and 299:
Page 300 and 301:
Page 302 and 303:
Page 304 and 305:
1568 I. J. Radiation Oncology d Bio
Page 306 and 307:
Page 308 and 309:
Page 310:
show all

LIFE01200604005 Shri Somnath Ghosh - Homi Bhabha National ...

You also want an ePaper? Increase the reach of your titles

Delete template?

Save as template?