Fig. 4: TG02 reduced Mcl-1 and induced apoptosis in the CLL cells at the presence of stroma protection.

A TG02 overcame stroma cell protection. CLL cells were cultured in RPMI media supplemented with 10% autologous plasma, either alone (media only, white bars), or co-culture with a layer of StromaNKtert cells (+stroma, black bars) overnight. Then the cells were incubated with increasing concentrations of TG02 and viabilities were analyzed by Annexin V/PI double staining after 4 and 24 h incubation (left), and Mcl-1 expression were quantitated by immunoblotting (right). Data present percentage of media only controls (mean ± SEM) of four individual CLL samples. B A representative immunoblot of cells incubated in conditions described in A. C Mcl-1 expression was induced by co-culturing with the StromaNKtert cells, which was reduced by TG02. CLL cells were cultured in RPMI media supplemented with 10% autologous plasma, either alone (media only, white bars), or co-culture with a layer of StromaNKtert cells (+stroma, black bars) overnight. Then the cells were incubated 1 μM TG02. Cell pellets were collected at 0, 1, 2, 4, and 6 h. Phosphorylation of RNA pol II and expression of Mcl-1 were analyzed by immunoblotting. Mcl-1 levels were quantitated in these samples and presented as levels relevant to media only control (left) and relevant to untreated controls (right) (mean ± SEM, n = 4). D A representative immunoblot of cells incubated in conditions described in (C).