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. 2013 Nov 12;8(11):e78443. doi: 10.1371/journal.pone.0078443

Figure 1. Simultaneous knockdown of HSPA1 and HSPA8 are necessary to compromise viability of MDA-MB-468 breast cancer cells.

Figure 1

MDA-MB-468 cells were transfected with siRNAs targeting the indicated heat shock proteins. For HSPA1 as well as HSPA8 and HSPA1+HSPA8 several single siRNAs or mixes were used marked with #1–3. A, Cell viability was measured six days after transfection by using Alamar Blue reagent. As positive control a pool of siRNAs against the mitotic kinase PLK1 was transfected. As negative controls non-targeting siRNAs (Co_4 and Co_5) were used, additionally cells were incubated only with the transfection reagent without siRNAs (TM) or left untreated (untr.). Viability is shown as % of TM. Only simultaneous transfection of HSPA1 and HSPA8 specific siRNAs induced a significant loss of viability. Additional knockdown of HSPA2 or HSPA5 had no further effect. B and C, Knockdown was confirmed by Western Blot analysis using HSPA1 and/or HSPA8 specific antibodies and infrared-labeled secondary antibodies three days after transfection of siRNAs. Knockdown efficiency was quantified by detecting signals with Odyssey infrared imaging system and is specified as % of TM.