Farnesyl transferase inhibitors induce extended remissions in transgenic mice with mature B cell lymphomas-3

D received transgenic lymphoma transplants 28 days previously (). The mice were left untreated or treated with L-744,832 for 28 days (L744 × 28) or for the final 7 days (L744 × 7). The numbers of B cells (upper left quadrant) and T cells (lower right quadrant) isolated from lymph nodes (first column), spleen (second column), or thymus (third column) were determined by flow cytometry after staining with antibodies to B220 and Thy1.2. The mean percentage and standard deviation of the B220and the Thy1.2cells are shown in the appropriate quadrant of each plot. There were 4 mice in all groups, except the L744 × 28 wild-type group, which had 2 mice.<p><b>Copyright information:</b></p><p>Taken from "Farnesyl transferase inhibitors induce extended remissions in transgenic mice with mature B cell lymphomas"</p><p>http://www.molecular-cancer.com/content/7/1/39</p><p>Molecular Cancer 2008;7():39-39.</p><p>Published online 19 May 2008</p><p>PMCID:PMC2409375.</p><p></p

Farnesyl transferase inhibitors induce extended remissions in transgenic mice with mature B cell lymphomas-5

S. The percent of viable splenocytes positive for both markers is shown for unmanipulated C57BL/6 mice (Wild-type) or mice transplanted with 10tumor cells 18 days earlier (Transplant recipient). Results from untreated mice are shown with black bars and from transplant-recipient mice treated with 1.56 mg SCH66336 by oral gavage twice daily for 3 days with shaded bars. The average and standard deviation for 4 mice in each group is shown.<p><b>Copyright information:</b></p><p>Taken from "Farnesyl transferase inhibitors induce extended remissions in transgenic mice with mature B cell lymphomas"</p><p>http://www.molecular-cancer.com/content/7/1/39</p><p>Molecular Cancer 2008;7():39-39.</p><p>Published online 19 May 2008</p><p>PMCID:PMC2409375.</p><p></p

Farnesyl transferase inhibitors induce extended remissions in transgenic mice with mature B cell lymphomas-2

Plenocytes are shown from unmanipulated C57BL/6 mice (Wild-type) or mice transplanted 18 days earlier with 10lymphoma cells from an Eμ-/BCR/HEL transgenic mouse (Transplant Recipient). Mice were either left untreated (black bars), treated orally for the last 3 days with 1.56 mg SCH66336 every 12 hours (shaded bars), or treated orally for the last 3 days with vehicle alone every 12 hours (clear bar). Each group contains 4 mice and error bars represent the standard deviation for each group.<p><b>Copyright information:</b></p><p>Taken from "Farnesyl transferase inhibitors induce extended remissions in transgenic mice with mature B cell lymphomas"</p><p>http://www.molecular-cancer.com/content/7/1/39</p><p>Molecular Cancer 2008;7():39-39.</p><p>Published online 19 May 2008</p><p>PMCID:PMC2409375.</p><p></p

Farnesyl transferase inhibitors induce extended remissions in transgenic mice with mature B cell lymphomas-1

Ild-type) or mice transplanted with 10Eμ-BCR/HEL transgenic lymphoma cells (Transplant recipient) were injected intravenously with 625 μg L-744,832 in 0.25 ml PBS daily (light bars) or left untreated (black bars). After 28 days, mice were euthanized and the spleens were dissected and viable cell counts of splenocytes resuspended in RPMI were determined using a hemocytometer. Error bars represent the standard deviation calculated for all mice in each group (n = 4). and . L-744,832 treatment causes rapid regression of lymphomas. In a separate experiment, mice transplanted with transgenic tumor cells were allowed to develop tumors for 20 or 24 days before daily treatment with L-744,832 as above for either 7 days (L744 × 7) or 3 days (L744 × 3), respectively. Twenty eight days following the tumor transplantation the mice were euthanized. Spleens were removed from tumor recipient mice and C57BL/6 control mice and photographed. Single cell suspensions from the spleens of mice that were left untreated (black bars), treated for the final 7 days (shaded bars), or treated for the final 3 days (clear bars) were counted using a hemocytometer. Data shown are the average of 4 mice in each group and error bars represent the standard deviation.<p><b>Copyright information:</b></p><p>Taken from "Farnesyl transferase inhibitors induce extended remissions in transgenic mice with mature B cell lymphomas"</p><p>http://www.molecular-cancer.com/content/7/1/39</p><p>Molecular Cancer 2008;7():39-39.</p><p>Published online 19 May 2008</p><p>PMCID:PMC2409375.</p><p></p

Farnesyl transferase inhibitors induce extended remissions in transgenic mice with mature B cell lymphomas-0

Ic mouse (panels -). The splenocytes were depleted of T cells and labeled with CFSE. Cells were cultured in RPMI (panels , -) or RPMI with 2 μg/ml anti-IgM and 1 μg/ml anti-CD40 (panels -) and 0 – 40 μM L-744,832 were included in the culture media, as indicated. After three days, cellular proliferation was measured by flow cytometry. Cells that have proliferated measure less than 10fluorescence units. For comparison, panels , and show proliferation in the absence of L-744,832 as a black line.<p><b>Copyright information:</b></p><p>Taken from "Farnesyl transferase inhibitors induce extended remissions in transgenic mice with mature B cell lymphomas"</p><p>http://www.molecular-cancer.com/content/7/1/39</p><p>Molecular Cancer 2008;7():39-39.</p><p>Published online 19 May 2008</p><p>PMCID:PMC2409375.</p><p></p

Farnesyl transferase inhibitors induce extended remissions in transgenic mice with mature B cell lymphomas-6

7 days with L-744,832 starting 21 days after transplantation (blue line) or left untreated (red line). In the first experiment (panel A), 5 mice each were treated with L-744,832 or left untreated. In the second experiment (panel B), 10 mice were treated with L-744,832 and 5 mice were untreated. Mice were then monitored for signs of lymphadenopathy and euthanized when moribund.<p><b>Copyright information:</b></p><p>Taken from "Farnesyl transferase inhibitors induce extended remissions in transgenic mice with mature B cell lymphomas"</p><p>http://www.molecular-cancer.com/content/7/1/39</p><p>Molecular Cancer 2008;7():39-39.</p><p>Published online 19 May 2008</p><p>PMCID:PMC2409375.</p><p></p

RNF11 is a negative regulator of virus-induced IFN-β production.

<p>(A) Micrographs of 293T cells transfected with either empty vector or Myc-RNF11 and then infected with VSV-GFP (MOI of 0.1) 24 h later. Pictures were taken 24 h post-infection. Immunoblotting was conducted with protein lysates using anti-Myc, anti-GFP and anti-Actin (right panel). (B) MEFs were transfected with either empty vector or Myc-RNF11 and were transfected again with poly(I:C) (15 μg) 24 h later. An IFN-β ELISA was performed 16 h later using supernatants. Immunoblotting was conducted with anti-Myc and anti-Actin. (C) 293T cells were transfected with an IFN-β luciferase reporter (200 ng), pRL-tk (20 ng), empty vector (1 µg) or RNF11-GFP (1 µg). Cells were transfected 24 h later with poly(I:C) (15 µg) and dual luciferase assays were performed after 16 h. Immunoblotting was conducted with protein lysates using anti-GFP and anti-Actin. (D) 293T cells were transfected with RNF11-GFP (1 μg) and Myc-A20 (1 μg) and then transfected 24 h later with poly(I:C) (20 µg). Immunoblotting was performed with anti-p-IRF3, anti-Myc, anti-GFP, anti-IRF3 and anti-Actin.</p

RNF11 interacts with TBK1/IKKi and blocks their Lys63-linked polyubiquitination.

<p>(A) 293T cells were transfected with 1 µg of RNF11-GFP, Flag-IKKi and Flag-TBK1. Co-IPs were conducted using anti-Flag for IP followed by immunoblotting with anti-GFP and anti-Flag. Immunoblotting was performed with lysates using anti-GFP, anti-Flag and anti-Actin. (B) 293T cells were transfected with poly(I:C) (20 μg) and co-IPs were performed with anti-RNF11 or isotype control IgG followed by immunoblotting with anti-IKKi (top panel) or anti-TBK1 (lower panel). Immunoblots were also performed with lysates using anti-IKKi, anti-RNF11, anti-TBK1 and anti-Actin. (C) 293T cells were transfected with empty vector, Flag-RNF11 or RNF11-GFP (1 µg) and HA-Ub-Lys63-only (500 ng). Cells were transfected again 24 h later with poly(I:C) (20 μg) and co-IPs were conducted the next day using anti-TBK1 (left panel) or anti-IKKi (right panel) followed by immunoblotting with anti-HA, anti-TBK1 (left panel) and anti-IKKi (right panel). Immunoblotting was performed with lysates with anti-Flag, anti-GFP and anti-Actin.</p

RNF11 inhibits IFN-β production at the level of TBK1/IKKi.

<p>(A–E) 293T cells were transfected with IFN-β luciferase reporter (200 ng), pRL-tk (20 ng), empty vector (1 μg), RNF11-GFP (1 μg) and either 0.5 μg of ΔRIG-I (A), MDA5 (B), IPS-1 (C), TBK1 (D) or IRF3-SA (E). Dual luciferase assays were performed with protein lysates 24 h later. (F, G) 293T cells were transfected with either control scrambled or RNF11 siRNA (60 pmol). After 24 h, cells were transfected with IFN-β luciferase reporter (200 ng), pRL-tk (20 ng), and either Flag-TBK1 or Flag-IKKi (0.5 μg) and dual luciferase assays were performed 24 h later. (H) 293T cells were transfected with either control scrambled or RNF11 siRNA (60 pmol). After 48 h, RT-PCR was performed to detect RNF11 and Actin transcripts. *, p<0.05. <i>Error bars, S.D.</i></p

Farnesyl transferase inhibitors induce extended remissions in transgenic mice with mature B cell lymphomas-7

Ic mouse (panels -). The splenocytes were depleted of T cells and labeled with CFSE. Cells were cultured in RPMI (panels , -) or RPMI with 2 μg/ml anti-IgM and 1 μg/ml anti-CD40 (panels -) and 0 – 40 μM L-744,832 were included in the culture media, as indicated. After three days, cellular proliferation was measured by flow cytometry. Cells that have proliferated measure less than 10fluorescence units. For comparison, panels , and show proliferation in the absence of L-744,832 as a black line.<p><b>Copyright information:</b></p><p>Taken from "Farnesyl transferase inhibitors induce extended remissions in transgenic mice with mature B cell lymphomas"</p><p>http://www.molecular-cancer.com/content/7/1/39</p><p>Molecular Cancer 2008;7():39-39.</p><p>Published online 19 May 2008</p><p>PMCID:PMC2409375.</p><p></p