Fig. 3.

TMEM205 mediates chemoresistance via the exosomal pathway and is overcome by combination oHSV+CP treatment. A Exosomes isolated from equal number of OVTOKO cells, OVTOKO cells with TMEM205 knocked down (OV TMSi), or OVTOKO cells with scrambled siRNA (Scr siRNA) control were quantified using NTA (n = 4, p0.05). B OVTOKO cells and OV TMSi cells were treated with CP; cell-lysates and exosomes were subjected to ICP-MS for analyzing CP accumulation (n = 3, p0.001). C JHOC and OVTOKO cells were either treated with CP alone or with either GW4869 or sphingomyelinase followed by CP. DNA extracted from the resulting cells was analyzed for CP DNA adduct by ICP MS. D SRB assay for OVTOKO cells with CP, oHSV and oHSV+CP. Percent survival was significantly lower in the combination group. Heat killed virus (attenuated oHSV) was used as a control (n = 6, p0.05, or 0.01). E Flowcytometry of oHSV and CP treated (GFP labeled) or only labeled cisplatin treated OV cells showing red cisplatin. F OVTOKO cells were treated with either CP or oHSV alone and the cell lysates were subjected to WB. TMEM205 and Akt were significantly reduced in the cells treated with oHSV. G Quantification (pixel density) of dot blot of genomic DNA isolated from OVTOKO cells treated with CP alone or oHSV+CP. H) DNA isolated from cells which were treated with CP or oHSV alone or the combination was run on an 0.8% agarose gel; H there was more DNA damage, in the form of shearing, with oHSV treatment (lane 1–8) and in the positive control where the DNA was treated with DNase (Lane1). Here Lane2 is DNA from JHOC cells, Lane 3 is from JHOC cells treated with heat killed oHSV, Lane 4 is from JHOC cells treated with heat killed oHSV followed by CP, Lane 5 is from JHOC cells treated CP, Lane 6 is from JHOC cells treated with oHSV, Lane 7 is from JHOC cells treated oHSV and CP, and Lane 8 is DNA treated with DNase