Figure S1. Rhod 2 Staining of Intracellular Calcium Depletion
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Figure S1. Rhod 2 staining of intracellular calcium depletion. Confocal microscopy bright field and fluorescent images x20 magnification of one UMM (OMM2.3) and one CMM (A375) line after staining with 1 µM Rhod 2 and 48 h of treatment with 0, 1, 3 and 8 µM amlodipine. UMM, uveal malignant melanoma; CMM, cutaneous malignant melanoma. Figure S2. Spheroid colonies. Light microscopy x5 magnification images of 3D spheroid colonies in CMM and UMM lines. Images were acquired just prior to the addition of 20 µM amlodipine in the treated groups or an equivalent volume of DMSO in control groups (Day 0). The same colonies were then visualized again after 48 h of treatment (Day 2). UMM, uveal malignant melanoma; CMM, cutaneous malignant melanoma. Figure S3. Senescence assay for amlodipine-induced cell death. Light microscopy x20 magnification images of CMM (A375, CHL-1 and IPC298) and UMM (MP41) lines when treated with amlodipine at 3 dose points over a 48-h period. Senescence is indicated with blue dye. UMM, uveal malignant melanoma; CMM, cutaneous malignant melanoma. Table SI. High-throughput drug screening compounds: Ranked list of the 1,018 screened compounds based on observed proliferation ratio. Rank Proliferation Compound ratio 1 0.00000 Fluoxetine HCl 2 0.00042 Amlodipine (Norvasc) 3 0.00042 Dronedarone HCl (Multaq) 4 0.00049 Prochlorperazine dimaleate 5 0.00055 Afatinib (BIBW2992) 7 0.00055 AMG-073 HCl (cinacalcet hydrochloride) 8 0.00055 Amlodipine besylate (Norvasc) 6 0.00055 Crizotinib (PF-02341066) 9 0.00064 Crystal violet 10 0.00074 Alexidine HCl 11 0.00074 Benzethonium chloride 14 0.00074 Cetrimonium bromide 13 0.00074 Domiphen bromide 15 0.00074 Thioridazine HCl 12 0.00074 Trifluoperazine 2HCl 16 0.00083 Bosutinib (SKI-606) 17 0.00085 Mitoxantrone hydrochloride 18 0.00085 Duloxetine HCl (Cymbalta) 19 0.00099 Cetylpyridinium chloride 20 0.00099 Thonzonium bromide 21 0.00106 Paroxetine HCl 22 0.00106 Carfilzomib (PR-171) 23 0.00110 Nebivolol (Bystolic) 24 0.00124 Nelfinavir mesylate 25 0.00124 Triflupromazine HCl 26 0.00127 Daunorubicin HCl (Daunomycin HCl) 27 0.00138 Doxorubicin (Adriamycin) 28 0.00149 Sertraline HCl 29 0.00169 Bazedoxifene HCl 30 0.00170 Loperamide hydrochloride 31 0.00173 Penfluridol 32 0.00193 Bortezomib (Velcade) 33 0.00197 9-Aminoacridine 34 0.00220 Epirubicin hydrochloride 35 0.00233 Aliskiren hemifumarate 37 0.00234 Chlorpromazine (Sonazine) 38 0.00234 Maprotiline hydrochloride 36 0.00234 Menadione 39 0.00247 Ouabain 40 0.00247 Sunitinib malate (Sutent) 41 0.00271 Cepharanthine 42 0.00271 Terfenadine 43 0.00321 Emetine 44 0.00519 Nitarsone 45 0.00568 Digoxigenin 46 0.00692 Clemastine fumarate 47 0.00741 Disulfiram (Antabuse) 48 0.01042 Clomipramine hydrochloride (Anafranil) 49 0.01048 Adefovir Dipivoxil (Preveon, Hepsera) 50 0.01212 Vinorelbine tartrate 51 0.01356 Proadifen HCl 52 0.01381 Clomifene citrate (Serophene) 53 0.01603 Thiostrepton 54 0.01762 Ethacridine lactate monohydrate 55 0.02190 Toremifene citrate (Fareston, Acapodene) 56 0.02229 Vorinostat (SAHA) 57 0.02435 Topotecan HCl 58 0.02618 Lomerizine HCl 59 0.02850 Ivacaftor (VX-770) 60 0.03878 Camptothecin 61 0.04406 Ziprasidone hydrochloride 62 0.04793 Sorafenib (Nexavar) 63 0.05525 Pyrithione zinc 64 0.05687 Ticagrelor 65 0.05819 Masitinib (AB1010) 66 0.09668 Meclocycline sulfosalicylate 67 0.11141 Asenapine 68 0.12470 Indacaterol maleate 69 0.13885 Carvedilol 70 0.13952 Primaquine diphosphate 71 0.13963 Bleomycin sulfate 72 0.15462 Imatinib Mesylate 73 0.15497 Solifenacin succinate 74 0.15876 Amitriptyline HCl 75 0.15924 R-(-)-Apomorphine HCl hemihydrate 76 0.18245 Pitavastatin calcium (Livalo) 77 0.20079 Ivermectin 78 0.20520 Desloratadine 79 0.20598 Pentamidine 80 0.20982 Aprepitant (MK-0869) 81 0.22060 Dapoxetine hydrochloride (Priligy) 82 0.22252 Cilnidipine 83 0.23689 Nifuroxazide 84 0.23974 Lonidamine 85 0.24045 Gemcitabine (Gemzar) 86 0.24876 Elvitegravir (GS-9137) 87 0.25162 Doxycycline HCl 88 0.25189 Cephalomannine 89 0.25404 Fenoprofen calcium 90 0.26206 Teniposide (Vumon) 91 0.26917 Chlorpheniramine maleate 92 0.27078 Tenofovir disoproxil Fumarate 93 0.29291 Riluzole (Rilutek) 94 0.29338 Mycophenolic (Mycophenolate) 95 0.29350 Evista (Raloxifene hydrochloride) 96 0.29821 XL-184 (Cabozantinib) 97 0.30870 Vinblastine 98 0.31176 Nisoldipine (Sular) 99 0.31183 Simvastatin (Zocor) 100 0.31240 Azacitidine (Vidaza) 101 0.32977 Pizotifen malate 102 0.33260 Albendazole oxide (Ricobendazole) 103 0.33269 Procodazole 104 0.33805 Azaguanine-8 105 0.34341 Lamivudine (Epivir) 106 0.34524 Ciclopirox (Penlac) 107 0.34591 Fluvoxamine maleate 108 0.35166 Clofarabine 109 0.35722 Valnemulin HCl 110 0.35954 Azelastine hydrochloride (Astelin) 111 0.36185 Licofelone 112 0.36509 Rotigotine 113 0.36512 Cladribine 114 0.36527 Darifenacin HBr 115 0.36631 Celecoxib 116 0.36805 Diperodon HCl 117 0.37690 L-Adrenaline (Epinephrine) 118 0.37842 Ciclopirox ethanolamine 119 0.38325 Nitazoxanide (Alinia, Annita) 120 0.38568 Acemetacin (Emflex) 121 0.39396 Trimipramine maleate 122 0.39694 Vincristine 123 0.40995 Coumarin 124 0.41325 Dasatinib (BMS-354825) 125 0.41573 Etravirine (TMC125) 126 0.41657 Doxazosin mesylate 127 0.42478 Cabazitaxel (Jevtana) 128 0.42821 Albendazole (Albenza) 129 0.43579 Chlorprothixene 130 0.43960 Lapatinib Ditosylate (Tykerb) 131 0.44312 Etoposide (VP-16) 132 0.44600 OSI-420 (Desmethyl erlotinib) 133 0.45329 Mycophenolate mofetil (CellCept) 134 0.45508 Miconazole nitrate 135 0.45794 Erlotinib HCl 136 0.46496 Pimobendan (Vetmedin) 137 0.47266 Lopinavir (ABT-378) 138 0.49230 Atorvastatin calcium (Lipitor) 139 0.49668 Domperidone (Motilium) 140 0.50606 Pazopanib HCl 141 0.50713 Clotrimazole (Canesten) 142 0.50750 Anisindione 143 0.50955 Fenbendazole (Panacur) 144 0.51170 Idarubicin HCl 145 0.51598 Benserazide 146 0.51712 Monobenzone (Benoquin) 147 0.51961 Irinotecan HCl trihydrate (Campto) 148 0.52007 Fludarabine (Fludara) 149 0.52252 Catharanthine 150 0.53186 Guanabenz acetate 151 0.53343 Nicardipine HCl 152 0.53592 Propafenone (Rytmonorm) 153 0.53665 Oxethazaine 154 0.53820 Deferasirox (Exjade) 155 0.54810 Fluvastatin sodium (Lescol) 156 0.55240 Carbidopa 157 0.55688 Tamoxifen citrate (Nolvadex) 158 0.56146 Clofoctol 159 0.56674 Fludarabine phosphate (Fludara) 160 0.57685 Imipramine HCl 161 0.57921 Irinotecan 162 0.58316 Resveratrol 163 0.58708 Chloroxine 164 0.58754 Trifluridine (Viroptic) 165 0.58785 Tacrine HCl 166 0.59079 2-Methoxyestradiol 167 0.59196 Tolcapone 168 0.59902 Econazole nitrate (Spectazole) 169 0.60221 Oxaliplatin (Eloxatin) 170 0.60308 Eltrombopag (SB-497115-GR) 171 0.60500 Paclitaxel (Taxol) 172 0.60974 Tigecycline 173 0.61596 Ethinyl Estradiol 174 0.61637 Brucine 175 0.61657 Benidipine hydrochloride 176 0.61796 10-DAB (10-Deacetylbaccatin) 177 0.61920 Ketoconazole 178 0.61991 Tolterodine tartrate (Detrol LA) 179 0.62391 Dicyclomine HCl 180 0.62849 Cyproheptadine HCl (Periactin) 181 0.62937 Phenazopyridine HCl 182 0.63254 Bufexamac 183 0.63641 Ulipristal 184 0.64024 Isoetharine mesylate 185 0.64187 Ospemifene 186 0.65023 Sulconazole nitrate 187 0.65218 Docetaxel (Taxotere) 188 0.65646 Oxytetracycline (Terramycin) 189 0.65812 Fenticonazole nitrate 190 0.66034 Ritonavir 191 0.66217 Benzydamine hydrochloride 192 0.66512 Imidapril (Tanatril) HCl 193 0.66684 Idoxuridine 194 0.66777 Miconazole (Monistat) 195 0.67937 Manidipine (Manyper) 196 0.68048 Axitinib 197 0.68169 Floxuridine 198 0.68281 Chlorquinaldol 199 0.70334 Cisatracurium besylate (Nimbex) 200 0.71341 Ivabradine HCl (Procoralan) 201 0.72356 Mesoridazine besylate 202 0.73867 Atazanavir sulfate 203 0.74086 Loratadine 204 0.74200 Nitrofurazone (Nitrofural) 205 0.74507 Imatinib (Gleevec) 206 0.74830 Fidaxomicin 207 0.75269 Hydroxyzine 2HCl 208 0.75730 Arbidol HCl 209 0.75825 Quinine hydrochloride dihydrate 210 0.75982 Dibucaine HCL 211 0.76361 Gefitinib (Iressa) 212 0.76433 Nifedipine (Adalat) 213 0.76709 Meticrane 214 0.76796 Atracurium besylate 215 0.76961 Deoxyarbutin 216 0.77569 Thiabendazole 217 0.78225 Phentolamine mesilate 218 0.78355 Formoterol hemifumarate 219 0.78515 Azelnidipine 220 0.78897 Progesterone (Prometrium) 221 0.79385 Tioconazole 222 0.79787 BIBR-1048 (Dabigatran) 223 0.79839 Naftopidil (Flivas) 224 0.79949 Exemestane 225 0.80059 Ezetimibe (Zetia) 226 0.80144 Fenoprofen calcium hydrate 227 0.80162 Methazolamide 228 0.80200 Rimonabant (SR141716) 229 0.80972 Quinapril HCl (Accupril) 230 0.81385 Regorafenib (BAY 73-4506) 231 0.82073 Flunarizine 2HCl 232 0.82128 Clozapine (Clozaril) 233 0.82618 Cobicistat (GS-9350) 234 0.82656 Curcumin 235 0.83127 Ranitidine (Zantac) 236 0.83376 Tolperisone HCl 237 0.83788 Mianserin hydrochloride 238 0.83835 Diphenylpyraline HCl 239 0.84359 Alprostadil (Caverject) 240 0.84467 Pramiracetam 241 0.85274 Broxyquinoline 242 0.85420 Estradiol 243 0.85457 Clorgyline HCl 244 0.85579 Benztropine mesylate 245 0.86009 Scopolamine hydrobromide 246 0.86178 Levosimendan 247 0.86179 Epinephrine bitartrate (Adrenalinium) 248 0.86669 Serotonin HCl 249 0.87069 Budesonide 250 0.87180 Alverine Citrate 251 0.87755 Rimantadine (Flumadine) 252 0.88126 Terbinafine hydrochloride (Lamisil) 253 0.88370 Scopine 254 0.88499 Carmofur 255 0.88530 Conivaptan HCl (Vaprisol) 256 0.88551 Mifepristone (Mifeprex) 257 0.88611 Tetracycline HCl 258 0.88968 Ethoxzolamide 259 0.89124 Noscapine HCl 260 0.89630 Methacycline hydrochloride (Physiomycine) 261 0.89960 Oxybuprocaine HCl 262 0.90202 Tiotropium bromide hydrate 263 0.90270 Triclabendazole 264 0.90285 Temsirolimus (Torisel) 265 0.90419 Adrucil (Fluorouracil) 266 0.90446 Nilvadipine (ARC029) 267