Fig 1.
Effect of LUX and IB on anti-IgM-induced BTK activation in SU-DHL-6 cells.
SU-DHL-6 cells were pre-treated for 2 h with either LUX or IB and then stimulated with anti-human IgM or left unstimulated. (A) Western blot analysis of whole cell lysates for p-BTK (Y551 and Y223), BTK, p-PLCγ2, PLCγ2 and GAPDH; representative blots are shown. (B-D) Quantification of intensity of bands in Western blots shown in A. Data is mean ± SEM from 2 independent experiments.
Fig 2.
Effect of LUX and IB on anti-IgM-induced activation of BLNK and BLNK-BTK interactions in SU-DHL-6 cells.
(A) Western blot analysis of whole cell lysates for p-BLNK (Y96), BLNK and GAPDH in SU-DHL-6 cells pre-treated for 2 h with either LUX or IB and then stimulated with anti-human IgM or left unstimulated. (B) Representative blots showing BLNK immunoprecipitates probed for BTK and BLNK in SU-DHL-6 cells sampled at 5, 10 and 15 min post anti-IgM stimulation. Data is mean ± SEM of data from 2 independent experiments.
Fig 3.
Effect of LUX and IB on anti-IgM-induced activation of SYK in SU-DHL-6 cells.
SU-DHL-6 cells were pre-treated for 2 h with either LUX or IB and then stimulated with anti-human IgM or left unstimulated. (A) Western blot analysis of whole cell lysates for p-SYK (Y525/Y526), SYK, and GAPDH; representative blots are shown. (B) Quantification of intensity of bands in Western blots shown in A. Data is mean ± SEM of data from 2 independent experiments.
Fig 4.
Effect of LUX and IB on LYN and CD79A in SU-DHL-6 cells.
SU-DHL-6 cells were pre-treated for 2 h with either LUX or IB and then stimulated with anti-human IgM or left unstimulated. (A) Western blot analysis of whole cell lysates for p-CD79A (Y182), CD79A, p-LYN (Y397)/LCK (Y394)/HCK (Y411)/BLK (Y389), LYN and GAPDH; representative blots are shown. (B-D) Quantification of intensity of bands in Western blots shown in A. Data is mean ± SEM of data from 2 independent experiments.
Fig 5.
Effect of LUX and IB on anti-IgM-induced phosphorylation of LYN in SU-DHL-6 cells.
Cells were pre-treated with vehicle or 1 μM LUX or IB after which they were stimulated with anti-human IgM. LYN immunoprecipitates from these cells were analyzed using antibodies for phospho-Src (Y416) and LYN at the indicated time points in SU-DHL-6 cells. Histograms show quantification of phospho-Src (Y416) band intensity relative to that of LYN. Data is mean ± SEM of data from 2 independent experiments. In the LUX + anti-IgM stimulated group, a statistically significant inhibition of p-Src (Y416) is denoted by @ (p<0.0001), # (p = 0.0003), a (p = 0.001), b (p = 0001) when compared to the respective time point for the anti-IgM stimulated cells. No significant inhibition of p-Src (Y416) was observed in the IB + anti-IgM group when compared to the respective time point for the anti-IgM stimulated cells.
Fig 6.
Effect of LUX and IB on IgM-induced phosphorylation of LYN (Y507) in SU-DHL-6, JeKo-1 and RL cells.
Cells were pre-treated for 2 h with either LUX or IB following which they were stimulated with anti-human IgM or left unstimulated. (A) Whole cell lysates were analyzed by Western blotting for phospho-LYN (Y507) and LYN; representative blots are shown. (B-D) Quantification of intensity of bands in Western blots shown in A. Data is mean ± SEM of data from 2 independent experiments.
Fig 7.
Summary of important kinases of the BCR signaling pathway whose phosphorylation is reduced by LUX following activation by anti-IgM in lymphoma cells.
The figure was created with Biorender.com.