Copyright information:Taken from "Differential expression and role of p21and p27in TNF-α-induced inhibition of proliferation in human glioma cells"http://www.molecular-cancer.com/content/6/1/42Molecular Cancer 2007;6():42-42.Published online 12 Jun 2007PMCID:PMC1904457.B) LN-229 cells were transfected with IκBα dominant negative construct and the expression of nuclear p65 was determined after exposure to TNF-α for 6 hr

Anjali Shiras (76149)

Gowry Das (76150)

Jayashree C Jagtap (76151)

Pabbisetty Sudheer Kumar (76148)

Padma Shastry (76153)

Vandna Prasad (76152)

Copyright information:Taken from "Differential expression and role of p21and p27in TNF-α-induced inhibition of proliferation in human glioma cells"http://www.molecular-cancer.com/content/6/1/42Molecular Cancer 2007;6():42-42.Published online 12 Jun 2007PMCID:PMC1904457.es (200 nM) for 6 hr and cultured as monolayers or spheroids for 18 hr. Cells were treated with TNF-α (10 ng/ml) for 24 hr, and tritiated thymidine incorporation assay was done as described in Fig. 1. * p < 0.05 inhibition of proliferation with TNF-α in transfected versus untransfected cells. The data is representative of three experiments done in triplicates

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Differential expression and role of p21and p27in TNF-α-induced inhibition of proliferation in human glioma cells-6

Copyright information:Taken from "Differential expression and role of p21and p27in TNF-α-induced inhibition of proliferation in human glioma cells"http://www.molecular-cancer.com/content/6/1/42Molecular Cancer 2007;6():42-42.Published online 12 Jun 2007PMCID:PMC1904457.or 24 hr, during the last 18 hr of treatment, 1 μCi of triatiated thymidine was added to the wells. The cells were harvested and the radio activity-counts per minute (CPM) was measured and percent inhibition of proliferation was calculated considering the CPM in untreated cells as 100%. B) Expression of TNFR1 and TNFR2 was analyzed by RT-PCR in LN-18 cells and compared with human glioma cell line-U87MG, used as positive control. β-actin was used as control. C) Expression of TNFR1 and TNFR2 in LN-18 cells and U87MG cells analyzed by flowcytometry, bold lines represent TNFR1/TNFR2 expression and dotted lines depict profiles with isocontrol antibody. D) Monolayers and spheroids were preincubated with neutralizing antibodies to TNFR1 and TNFR2 (50 μg/ml) or isocontrol antibody for 2 hr and treated with TNF-α (10 ng/ml) for 24 hr and proliferation assay was done as described above. E) Monolayers and spheroids were treated with serial concentrations of TNFR1 agonistic antibody or isocontrol antibody for 24 hr and proliferation was determined by triatiated thymidine incorporation assay. The data for A, D and E is the mean ± SE of at least three independent experiments done in triplicates. * p < 0.05-comparison of inhibition of proliferation in TNF-α treated cells in the presence and absence of TNFR neutralizing antibodies for D and between treated and controls cells for A and E. F) Levels of TNFR1 expression by Western blotting in monolayers (M) and spheroids (S) cultured for 24 hr. The blots were stripped and reprobed with β-actin, which served as a loading control of proteins

Differential expression and role of p21and p27in TNF-α-induced inhibition of proliferation in human glioma cells-0

Differential expression and role of p21and p27in TNF-α-induced inhibition of proliferation in human glioma cells-3

Differential expression and role of p21and p27in TNF-α-induced inhibition of proliferation in human glioma cells-8

Copyright information:Taken from "Differential expression and role of p21and p27in TNF-α-induced inhibition of proliferation in human glioma cells"http://www.molecular-cancer.com/content/6/1/42Molecular Cancer 2007;6():42-42.Published online 12 Jun 2007PMCID:PMC1904457.done by flowcytometry. The percent population of cells in G0/G1, S and G2/M phases of cell cycle was determined using Modfit software. The plot is a representative of three similar experiments. B) Monolayers (M) and spheroids(S) were treated with TNF-α (10 ng/ml) for 3 hr and the expression of p21 and p27 was determined by Western blotting. Blots were reprobed for β-actin and used as control for equal loading of proteins. . LN-18 cells were grown on coverslips for 24 hr and treated with TNF-α (10 ng/ml) for different time periods. Cells were stained with antibody recognizing nuclear and cytoplasmic p21, or with p27 antibody followed by secondary antibody labeled with Cy3. The expression of p21/p27 is depicted as red fluorescence. DAPI was used for nuclear staining (blue). The merged images show nuclear localization of p21 and p27 localized predominantly in the cytoplasm. (Magnification 63×)

Differential expression and role of p21and p27in TNF-α-induced inhibition of proliferation in human glioma cells-1

Copyright information:Taken from "Differential expression and role of p21and p27in TNF-α-induced inhibition of proliferation in human glioma cells"http://www.molecular-cancer.com/content/6/1/42Molecular Cancer 2007;6():42-42.Published online 12 Jun 2007PMCID:PMC1904457.ed by MTT assay. The percent cell death was determined considering viability of control cells as 100%. (B) IκBα mutant LN-18 cells were treated with TNF-α (10 ng/ml) for different time points and cleavage of PARP (89 kD fragment) was determined by Western blotting. C) IκBα mutant LN-18 cells grown on coverslips were treated with TNF-α (10 ng/ml) for 3 hr and stained with p21 followed by secondary antibody labeled with Cy3 and DAPI was used for staining nucleus (Magnification 63×). D) Expression of p21 and p27 determined by Western blotting in monolayers (M) and spheroids (S) derived from IκBα mutant LN-18 cells treated with TNF-α for 3 hr. The blots were stripped and reprobed with β-actin, which served as control for equal loading of proteins

Differential expression and role of p21and p27in TNF-α-induced inhibition of proliferation in human glioma cells-2

https://figshare.com/articles/Differential_expression_and_role_of_p21and_p27in_TNF_induced_inhibition_of_proliferation_in_human_glioma_cells_0/58064

Differential expression and role of p21and p27in TNF-α-induced inhibition of proliferation in human glioma cells-3

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