Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Mitochondrial dysfunction induced by a SH2 domain-targeting STAT3 inhibitor leads to metabolic synthetic lethality in cancer cells

doi: 10.1073/pnas.1615730114

Figure Lengend Snippet: OPB-51602 impairs mitochondrial function. (A) MMP (JC1 staining) in DU145 cells treated with OPB-51602 and OPB-31121 for 2 h in CM. (B) Mitochondrial OCR in control and OPB-51602–treated (100 nM for 2 h) DU145 cells. (C) Mitochondrial shape in DU145 cells treated with OPB-51602 (100 nM) in CM (16 h) or glucose-depleted medium (4 h) and stained with MitoTracker Orange (red) and DAPI (blue). (D) MMP (DIOC6 staining) in isolated mitochondria from DU145 cells treated with OPB-51602 (50 μM) or Valinomycin (100 μM) for 3 h. (E) MMP (DIOC6 staining) in isolated mitochondria from WT and STAT3−/− MEF treated with OPB-51602 for 3 h. (F) Cell viability [sulforhodamine B (SRB) assay] in DU145 and DU145 ρ° cells treated with OPB-51602. *P < 0.01.

Article Snippet: After 48 h, cells were treated with the drugs for 6 h, then fixed with 4% paraformaldehyde, and mounted with VECTASHIELD Mounting Medium with DAPI (Reactolab).

Techniques: Staining, Control, Isolation, Sulforhodamine B Assay

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Mitochondrial dysfunction induced by a SH2 domain-targeting STAT3 inhibitor leads to metabolic synthetic lethality in cancer cells

doi: 10.1073/pnas.1615730114

Figure Lengend Snippet: Induction of STAT3 protein aggregates by OPB-51602. (A) Distribution of EGFP-STAT3 (green) in DU145 and H358 cells untreated or treated with OPB-51602 for 6 h in CM. Nuclei were stained with DAPI (blue). (B) Effect of OPB-51602 on WT and mutated (E638A) EGFP-STAT3. (C and D) WT and truncated variants of EGFP-STAT3 (C) and differential responses to treatment with OPB-51602 (D). (E) Spontaneous aggregation of ΔSH2 EGFP-STAT3 variant in CM cultures. (F) Live fluorescence, phase contrast, and composite images of H358 cells expressing WT or ΔSH2 EGFP-STAT3 at the indicated time points. Arrows indicate the position of EGFP+ cells in phase contrast images.

Article Snippet: After 48 h, cells were treated with the drugs for 6 h, then fixed with 4% paraformaldehyde, and mounted with VECTASHIELD Mounting Medium with DAPI (Reactolab).

Techniques: Staining, Variant Assay, Fluorescence, Expressing