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Table 1 Plasmids used

From: Protease 3C of hepatitis A virus induces vacuolization of lysosomal/endosomal organelles and caspase-independent cell death

Plasmid Description Reference/Source
Vectors encoding organelle-targeted fluorescent proteins
pmKate2-lyso encodes mKate2 N-terminally fused to rat Lamp-1 Evrogen (Russia)
pTagCFP-mito encodes TagCFP N-terminally fused to mitochondrial targeting sequence derived from the subunit VIII of human cytochrome C oxidase Evrogen (Russia)
pTagRFP-Golgi encodes TagRFP N-terminally fused to Golgi targeting sequence of human β-1,4-galactosyltransferase Evrogen (Russia)
pRab7-EYFPa encodes Rab7 C-terminally fused to eYFP [88]
pRab5-ECFPa encodes Rab5 C-terminally fused to eCFP [88]
pDsRed-Rab5/DNb encodes dominant-negative Rab5/N133I N-terminally fused to DsRed [134,135]/Add #13051g
pDsRed-Rab7/DNb encodes Rab7/T22N N-terminally fused to DsRed [134,135]/Add #12662g
pDsRed-Rab9b encodes Rab9 N-terminally fused to DsRed [134,135]/Add #12677g
pDsRed-Rab11b encodes Rab11 N-terminally fused to DsRed [134,135]/Add #12679g
pmRFP-LC3c encodes autophagosome-specific LC3 protein N-terminally fused to mRFP [136]/Add #21075g
pYFP-Rac1(N17)d encodes Rac1/T17N N-terminally fused to fluorescent protein YFP [137]/Add #11395g
pER-RFPe encodes RFP N-terminally fused to CD5 leader sequence and C-terminally fused to SEKDEL amino acid sequence [43]
Vectors for tet-off advanced expression system
pTet-Off Advanced encodes the tetracycline-controlled transactivator protein tTA-Advanced Clontech (USA)
pBI-EGFP allows expression of a gene of interest and marker gene of eGFP under control of the bi-directional tTA-responsive promotor Clontech (USA)
3Cpro gene source
pHAV3’f bears a cDNA encoding a segment of human hepatitis A virus genome (strain HAS-15) [138]/GenBank: X15463.1
3Cpro-expressing plasmids
pBI-EGFP/3C encodes 3Cpro and eGFP under control of bi-directed tTA-responsive promotor. This study
pBI-EGFP/3CMut encodes 3Cpro with Cys172→Ala mutation and eGFP under control of bi-directed tTA-responsive promotor. This study
  1. aGift from Prof. Ari Helenius (Institute of Biochemistry, Swiss Federal Institute of Technology, Zurich, Switzerland).
  2. bGift from Dr. Richard Pagano (Mayo Clinic, Rochester, MN, USA).
  3. cGift from Dr. Tamotsu Yoshimori (National Institute for Basic Biology, Okazaki, Japan).
  4. dGift from Dr. Joel Swanson (University of Michigan Medical School, MI, USA).
  5. eGift from Dr. Felipe X. Pimentel-Muiños (Centro de Investigation del Cancer, Universidad de Salamanca-CSIC, Salamanca, Spain).
  6. fGift from Dr. Eugene Snezhkov (M.M. Shemyakin and Yu.A Ovchinnikov Institute of Bioorganic Chemistry RAS, Moscow, Russia).
  7. gPlasmid was distributed through Addgene. Add #, Addgene plasmid number.