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. 2016 Mar 16;11(3):e0150044. doi: 10.1371/journal.pone.0150044

Fig 5. ASS1 knockdown by siRNA increases the chemosensitivity of spheroids.

Fig 5

(A) Immunoblot analysis of M28, SARC, VAMT and ASS1-deficient MSTO spheroids, transfected with siRNA targeting ASS1 or control sequences show that ASS1 was efficiently knocked down in all the cells where it was expressed. (B) Apoptotic rates were measured for M28, SARC, VAMT and MSTO spheroids transfected with siRNA targeting ASS1 or control sequences and treated with bortezomib (BZ, 25 nM) or cis-platinum (200 μM) plus pemetrexed (10 μM)(C+P). In the 3 cell lines with baseline ASS1 expression, ASS1 siRNA increased the basal apoptotic rate and also increased the apoptotic response to chemotherapy. ASS1 siRNA had no effect in the ASS1-deficient MSTO spheroids. Apoptosis was quantified in a blinded fashion by assessing nuclear condensation in Hoechst-stained cells. (* p ≤ 0.05 n = 3).