p38α  (MedChemExpress)

Journal: International Journal of Molecular Sciences

Article Title: Perturbation of p38α MAPK as a Novel Strategy to Effectively Sensitize Chronic Myeloid Leukemia Cells to Therapeutic BCR-ABL Inhibitors

doi: 10.3390/ijms222212573

Figure Lengend Snippet: Knockdown of p38α had no apparent effects on the viability of BCR-ABL-positive K562 leukemia cells. The transcripts of p38α in K562 cells were knocked down using specific shRNAs. The protein levels of p38α were examined by Western blotting ( A ). The cell growth of parental K562 and knockdown KD1 cell clones were examined using hemocytometer counting under a microscope ( B ). The viable ( C ) and dead cells ( D ) were identified by trypan blue exclusion. All results shown are representatives of three independent experiments. Cell number and viability are presented as the mean ± SE of three repeats. * p < 0.05, and *** p < 0.005.

Article Snippet: To test the effect of specific p38α inhibitor, TAK715 (10 mM stock in DMSO) (HY-10456, MedChemExpress, Monmouth Junction, NJ, USA) was added into cells 1 h prior to the addition of imatinib or dasatinib.

Techniques: Knockdown, Western Blot, Clone Assay, Microscopy

Journal: International Journal of Molecular Sciences

Article Title: Perturbation of p38α MAPK as a Novel Strategy to Effectively Sensitize Chronic Myeloid Leukemia Cells to Therapeutic BCR-ABL Inhibitors

doi: 10.3390/ijms222212573

Figure Lengend Snippet: Knockdown of p38α significantly enhanced imatinib-induced cytotoxicity in BCR-ABL-positive K562 leukemia cells. The K562 parental and p38α-knockdown (KD1) cells were treated with 0.3 µM ( A ), 0.6 µM ( B ), and 1.2 µM ( C ) of imatinib. The viability of cells was examined by trypan blue exclusion. Knockdown of p38α greatly enhanced the therapeutic efficacy of imatinib. All results shown are representatives of three independent experiments. Total and dead cell numbers are presented as the mean ± SE of three repeats. * p < 0.05, ** p < 0.01, and *** p < 0.005.

Article Snippet: To test the effect of specific p38α inhibitor, TAK715 (10 mM stock in DMSO) (HY-10456, MedChemExpress, Monmouth Junction, NJ, USA) was added into cells 1 h prior to the addition of imatinib or dasatinib.

Techniques: Knockdown

Journal: International Journal of Molecular Sciences

Article Title: Perturbation of p38α MAPK as a Novel Strategy to Effectively Sensitize Chronic Myeloid Leukemia Cells to Therapeutic BCR-ABL Inhibitors

doi: 10.3390/ijms222212573

Figure Lengend Snippet: Imatinib-induced apoptotic cells death significantly increased when p38α was deficient. The K562 parental and p38α-knockdown (KD1) cells were treated with various concentrations of imatinib as indicated for 48 and 72 h. Cells were fixed and stained with propidium iodide. The cell cycle distribution was analyzed by flow cytometry: ( A ) 48 h and ( B ) 72 h. Knockdown of p38α significantly increased the apoptotic sub-G1 population after imatinib treatment. Activation of caspase-3 was analyzed by Western blotting using specific antibodies ( C ). Etoposide (Ep) is a chemotherapeutic drug known to induce apoptosis and used as a positive control. The ratio of band intensity was quantified as described in . Both the active form of caspase-3 and the cleavage of the specific substrate PARP increased in p38α knockdown cells. All results shown are representatives of three independent experiments.

Article Snippet: To test the effect of specific p38α inhibitor, TAK715 (10 mM stock in DMSO) (HY-10456, MedChemExpress, Monmouth Junction, NJ, USA) was added into cells 1 h prior to the addition of imatinib or dasatinib.

Techniques: Knockdown, Staining, Flow Cytometry, Activation Assay, Western Blot, Positive Control

Journal: International Journal of Molecular Sciences

Article Title: Perturbation of p38α MAPK as a Novel Strategy to Effectively Sensitize Chronic Myeloid Leukemia Cells to Therapeutic BCR-ABL Inhibitors

doi: 10.3390/ijms222212573

Figure Lengend Snippet: Knockdown of p38β did not sensitize K562 leukemia cells to the therapeutic effects of imatinib. The transcripts of p38β were knocked down using specific shRNAs. The protein levels of p38β were detected by Western blotting ( A ). The cell growth and viability of parental K562 and knockdown KD1 cell clones under normal culturing conditions were examined by trypan blue exclusion using hemocytometer counting under a microscope ( A ). Cells were treated with 0.3 ( B ), 0.6 ( C ), and 1.2 µM ( D ) of imatinib and the cell viability was examined. Deficiency of p38β did not sensitize K562 cells to the killing effect of imatinib as observed with p38α deficiency. On the contrary, p38β deficiency exhibited a slight but significant resistance to the killing effect of imatinib. All results shown are representatives of three independent experiments. Viable and dead cell numbers are presented as the mean ± SE of three repeats. * p < 0.05, ** p < 0.01, and *** p < 0.005.

Article Snippet: To test the effect of specific p38α inhibitor, TAK715 (10 mM stock in DMSO) (HY-10456, MedChemExpress, Monmouth Junction, NJ, USA) was added into cells 1 h prior to the addition of imatinib or dasatinib.

Techniques: Knockdown, Western Blot, Clone Assay, Microscopy

Journal: International Journal of Molecular Sciences

Article Title: Perturbation of p38α MAPK as a Novel Strategy to Effectively Sensitize Chronic Myeloid Leukemia Cells to Therapeutic BCR-ABL Inhibitors

doi: 10.3390/ijms222212573

Figure Lengend Snippet: The cytotoxic effect of dasatinib was also enhanced upon p38α knockdown. The K562 parental and p38α-knockdown (KD11) cells were treated with 1 ( A ) or 2 nM ( B ) of the second-generation TKI dasatinib. The viability of cells was examined by trypan blue exclusion. Knockdown of p38α greatly enhanced the therapeutic efficacy of dasatinib. All results shown are representative of three independent experiments. Viable and dead cell numbers are presented as the mean ± SE of three repeats. ** p < 0.01, and *** p < 0.005.

Article Snippet: To test the effect of specific p38α inhibitor, TAK715 (10 mM stock in DMSO) (HY-10456, MedChemExpress, Monmouth Junction, NJ, USA) was added into cells 1 h prior to the addition of imatinib or dasatinib.

Techniques: Knockdown

Journal: International Journal of Molecular Sciences

Article Title: Perturbation of p38α MAPK as a Novel Strategy to Effectively Sensitize Chronic Myeloid Leukemia Cells to Therapeutic BCR-ABL Inhibitors

doi: 10.3390/ijms222212573

Figure Lengend Snippet: The p38α deficiency increased cellular sensitivity to imatinib in BCR-ABL-positive KBM5 leukemia cells. The p38α transcripts in KBM5 cells were knocked down using specific shRNAs. The protein levels of p38α were examined by Western blotting ( A ). The cell growth and viability of parental KBM5 and knockdown M5-8H and M5-11G cell clones were examined using hemocytometer counting under a microscope ( A ). Dead cells were identified by trypan blue exclusion. Cells were treated with 0.2 ( B ) and 0.5 µM ( C ) imatinib and the cell viability was examined. All results shown are representative of three independent experiments. Cell number and viability are presented as the mean ± SE of three repeats. * p < 0.05, ** p < 0.01, and *** p < 0.005.

Article Snippet: To test the effect of specific p38α inhibitor, TAK715 (10 mM stock in DMSO) (HY-10456, MedChemExpress, Monmouth Junction, NJ, USA) was added into cells 1 h prior to the addition of imatinib or dasatinib.

Techniques: Western Blot, Knockdown, Clone Assay, Microscopy

Journal: International Journal of Molecular Sciences

Article Title: Perturbation of p38α MAPK as a Novel Strategy to Effectively Sensitize Chronic Myeloid Leukemia Cells to Therapeutic BCR-ABL Inhibitors

doi: 10.3390/ijms222212573

Figure Lengend Snippet: The p38α deficiency increased cellular sensitivity to dasatinib in BCR-ABL-positive KBM5 leukemia cells. The KBM5 parental and p38α-knockdown (M5-11G and M5-8H) cells were treated with 0.5 ( A ) or 1 nM ( B ) of second-generation TKI dasatinib. The viability of cells was examined by trypan blue exclusion. Knockdown of p38α greatly enhanced the therapeutic efficacy of dasatinib. All results shown are representative of three independent experiments. Viable and dead cell numbers are presented as the mean ± SE of three repeats. * p < 0.05, ** p < 0.01, and *** p < 0.005.

Article Snippet: To test the effect of specific p38α inhibitor, TAK715 (10 mM stock in DMSO) (HY-10456, MedChemExpress, Monmouth Junction, NJ, USA) was added into cells 1 h prior to the addition of imatinib or dasatinib.

Techniques: Knockdown

Journal: International Journal of Molecular Sciences

Article Title: Perturbation of p38α MAPK as a Novel Strategy to Effectively Sensitize Chronic Myeloid Leukemia Cells to Therapeutic BCR-ABL Inhibitors

doi: 10.3390/ijms222212573

Figure Lengend Snippet: Specific p38α inhibitor TAK715 greatly increased the therapeutic efficacy of imatinib and dasatinib toward CML cells. K562 leukemia cells were treated with imatinib (0.3 µM) ( A ) or dasatinib (1 nM) ( B ) in combination with p38α inhibitor TAK715 (5 or 10 µM). Alternatively, cells were treated with TAK715 (5 or 10 µM) alone ( C ). The viability of cells was examined by trypan blue exclusion. All results shown are representative of three independent experiments. Viable and dead cell numbers are presented as the mean ± SE of three repeats. * p < 0.05, ** p < 0.01, and *** p < 0.005.

Article Snippet: To test the effect of specific p38α inhibitor, TAK715 (10 mM stock in DMSO) (HY-10456, MedChemExpress, Monmouth Junction, NJ, USA) was added into cells 1 h prior to the addition of imatinib or dasatinib.

Techniques: