(a): Interdependency of TIMP-1 and Bcl-2 proteins: H2009 cells and its clones were treated with 0.5μM Staurosporine (S), which showed a reduction of TIMP-1 protein expression. Treatment with a BH3 mimetic ABT-737(A) reduced TIMP-1 protein levels compared to the controls. Combined treatment with ABT-737 and Staurosporine (AS) showed a marked reduction of TIMP-1 expression, without any changes in TIMP-2. (b) TIMP-1 mRNA expression level after treatment with Staurosporine and ABT-737: HB1 and HB6 TIMP-1 levels were reduced with combined ABT-737 and Staurosporine treatment. Two Way Repeated Measure ANOVA showed p = 0.0063, indicating a very significant down regulation of relative expression of TIMP1 in the overexpressed clones after ABT+Staurosporine treatment indicating down-regulating Bcl2 affects TIMP1 level. (c) Co-immunoprecipitation of TIMP-1 and Bcl-2: A possible interaction between the Bcl-2 and TIMP-1 revealed by co-immunoprecipitation. Cell extracts from H2009 and it’s clones wer Srilatha Nalluri Sampa Ghoshal-Gupta Ammar Kutiyanawalla Sitaram Gayatri Byung Rho Lee Shahanawaz Jiwani Amyn M. Rojiani Mumtaz V. Rojiani 10.1371/journal.pone.0137673.g003 https://plos.figshare.com/articles/figure/_a_Interdependency_of_TIMP_1_and_Bcl_2_proteins_H2009_cells_and_its_clones_were_treated_with_0_5_956_M_Staurosporine_S_which_showed_a_reduction_of_TIMP_1_protein_expression_Treatment_with_a_BH3_mimetic_ABT_737_A_reduced_TIMP_1_protein_levels_compared_to_t/1541916 <p>(a): Interdependency of TIMP-1 and Bcl-2 proteins: H2009 cells and its clones were treated with 0.5μM Staurosporine (S), which showed a reduction of TIMP-1 protein expression. Treatment with a BH3 mimetic ABT-737(A) reduced TIMP-1 protein levels compared to the controls. Combined treatment with ABT-737 and Staurosporine (AS) showed a marked reduction of TIMP-1 expression, without any changes in TIMP-2. (b) TIMP-1 mRNA expression level after treatment with Staurosporine and ABT-737: HB1 and HB6 TIMP-1 levels were reduced with combined ABT-737 and Staurosporine treatment. Two Way Repeated Measure ANOVA showed p = 0.0063, indicating a very significant down regulation of relative expression of TIMP1 in the overexpressed clones after ABT+Staurosporine treatment indicating down-regulating Bcl2 affects TIMP1 level. (c) Co-immunoprecipitation of TIMP-1 and Bcl-2: A possible interaction between the Bcl-2 and TIMP-1 revealed by co-immunoprecipitation. Cell extracts from H2009 and it’s clones were immunoprecipitated (IP) with anti-Bcl-2 or normal rabbit IgG as a negative control. The immunoblot (IB) was probed for TIMP-1, TIMP-2 and Bcl-2. No TIMP-2 was pulled down with Bcl-2, showing that only TIMP-1 is associated with Bcl-2 when apoptosis is induced. Input represents 60μg of cell extract.</p> 2015-09-14 03:33:52 MMP inhibition activity overexpressing H 2009 cells protein levels hb Overexpressing TIMP cell lysates exhibit properties Inhibits Apoptosis TIMP 1 overexpressing clones PARP cleavage tumor kinetics T 2G Bax levels H 2009 lung adenocarcinoma cells lung cancer cell lines staurosporine treatment bad Cancer Biomarker rna p 90RSK lung adenocarcinoma cell line H 2009 bcl cns mitochondrial permeability lung adenocarcinoma cells