American Society of Hematology

Figure 2.

$Figure 2. NFκB activity and inhibition in an MLBCL cell line. (A) NFκB DNA-binding activity of the MLBCL cell line, Karpas 1106; 2 DLBCL cell lines, OCI LY10 and DHL6; and a Hodgkin lymphoma cell line, KM-H2. NFκB DNA-binding activity was measured using a colorimetric assay. Measurements were performed in triplicates. (B) NFκB activity and apoptosis in MLBCLs expressing an IκBα superrepressor. (Left) NFκB DNA-binding activity in MLBCL cells transduced with MSCV-eGFP (vector) alone or MSCV-eGFP-SR-IκBα. Measurements were performed in triplicate. NFκB activity was significantly lower in SR-IκBα cells than in vector-only cells (P < .001, one-sided Student t test). (Right) The apoptotic fraction (percentage annexin V+ GFP+) of cells transduced with MSCV-eGFP-SR-IκBα is much higher than that of cells transduced with vector alone. (C) Apoptosis in MLBCL cells expressing an IκB superrepressor. GFP-positive cells expressing vector alone or MSCV-eGFP-SR-IκBα were analyzed for expression of annexin V (Alexa 568). GFP, x-axis; annexin V, y-axis. (D) Proliferation of MLBCL cells expressing an IκBα superrepressor. The proliferation (MTS absorbance) of parental and GFP+ vector-only- and MSCV-eGFP-SR-IκBα-transduced MLBCL cells was measured at 0, 24, 48, and 72 hours and assessed with an ANOVA model that included the type of treatment and time of measurement. The difference between SR-IκBα-transduced MLBCL cells and either vector-only or parental cells was significant (P < .001 in each case), whereas there was no difference between empty vector and parental cells (P = NS). The measurements were performed in triplicate. All experiments (A-D) were performed 3 to 4 times with comparable results; representative experiments are shown. Error bars indicate the standard deviation within triplicate experiments.$

NFκB activity and inhibition in an MLBCL cell line. (A) NFκB DNA-binding activity of the MLBCL cell line, Karpas 1106; 2 DLBCL cell lines, OCI LY10 and DHL6; and a Hodgkin lymphoma cell line, KM-H2. NFκB DNA-binding activity was measured using a colorimetric assay. Measurements were performed in triplicates. (B) NFκB activity and apoptosis in MLBCLs expressing an IκBα superrepressor. (Left) NFκB DNA-binding activity in MLBCL cells transduced with MSCV-eGFP (vector) alone or MSCV-eGFP-SR-IκBα. Measurements were performed in triplicate. NFκB activity was significantly lower in SR-IκBα cells than in vector-only cells (P < .001, one-sided Student t test). (Right) The apoptotic fraction (percentage annexin V⁺ GFP⁺) of cells transduced with MSCV-eGFP-SR-IκBα is much higher than that of cells transduced with vector alone. (C) Apoptosis in MLBCL cells expressing an IκB superrepressor. GFP-positive cells expressing vector alone or MSCV-eGFP-SR-IκBα were analyzed for expression of annexin V (Alexa 568). GFP, x-axis; annexin V, y-axis. (D) Proliferation of MLBCL cells expressing an IκBα superrepressor. The proliferation (MTS absorbance) of parental and GFP⁺ vector-only- and MSCV-eGFP-SR-IκBα-transduced MLBCL cells was measured at 0, 24, 48, and 72 hours and assessed with an ANOVA model that included the type of treatment and time of measurement. The difference between SR-IκBα-transduced MLBCL cells and either vector-only or parental cells was significant (P < .001 in each case), whereas there was no difference between empty vector and parental cells (P = NS). The measurements were performed in triplicate. All experiments (A-D) were performed 3 to 4 times with comparable results; representative experiments are shown. Error bars indicate the standard deviation within triplicate experiments.

This Feature Is Available To Subscribers Only