Figure 3.

Manipulations in the protein levels of HSPA1 and HSPA2 have limited effect on sensitivity of NSCLC cells to manumycin A (MA). (a,c) Immunoblots showing levels of HSPA1 and HSPA2 in wild-type (wt) and lentivirally-modified cells; sh-luc control cells were transduced with a non-targeting shRNA-luc sequence; sh-A1.N and sh-A1.S cell lines were transduced with HSPA1-targeting shRNA sequences; control p-LVX cells were transduced with lentiviruses bearing the “empty” pLVX-Puro vector; p-A2 cells were transduced with pLVX-Puro plasmid encoding HSPA2 protein under the control of the CMV promoter; sh-A2.3 and sh-A2.4 cell lines were transduced with HSPA2-targeting shRNA sequences. Representative immunoblots are shown (n ≥ 3); actin was used as a protein loading control. These model cell lines were described in detail previously [26,27]. (b,d) Cell viability was measured using MTS assay after 72 h treatment with MA (0–10 µM). Results are expressed as mean ± SD in relation to untreated control (n = 4, each in three technical replicates, * p < 0.05, statistical significance was determined by two-tailed t-test). The numbers on the right side of immunoblots indicate molecular weight of the protein size marker.