Journal: Cell communication and signaling : CCS

Article Title: BMX, a specific HDAC8 inhibitor, with TMZ for advanced CRC therapy: a novel synergic effect to elicit p53-, β-catenin- and MGMT-dependent apoptotic cell death.

doi: 10.1186/s12964-022-01007-x

Figure Lengend Snippet: Fig. 1 BMX, TMZ, oxaliplatin (Oxp) and doxorubicin (Dox) combination inhibited cell proliferation in CRC cells. (A) The proliferation of BMX, TMZ, Oxp, Dox, BMX plus TMZ, BMX plus Oxp or BMX plus Dox in HT29, HCT116 and RKO cells with various drug concentration. (B) Colony formation capability assay with different treatments of BMX, TMZ, Oxp, BMX plus TMZ, and BMX plus Oxp in HT29, HCT116 and RKO cells; the colonies were counted for quantification. (C) Cell cycle analysis after 48 h treatment with different concentrations of BMX alone or BMX combined with TMZ in HT29, HCT116, and RKO cells and the proportion of cells in each cell cycle phase. SubG1, cell with polyploid chromosome; > 4 N, polyploid cell. (D) Apoptosis analysis after 48 h treatment with different concentrations of BMX alone or BMX combined with TMZ and the apoptotic rate of cells in HT29, HCT116, and RKO cells. All results are shown as mean ± s.d. from three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 versus control (HT29 cells); #p < 0.05, ##p < 0.01, ###p < 0.001 vs. control (HCT116 cells); †p < 0.05, ††p < 0.01, †††p < 0.001 versus control (RKO cells)

Article Snippet: The cell proliferation of CRC cells was measured by CCK8 assay (Targetmol, Shanghai, China).

Techniques: Concentration Assay, Cell Cycle Assay, Control

Journal: Cell communication and signaling : CCS

Article Title: BMX, a specific HDAC8 inhibitor, with TMZ for advanced CRC therapy: a novel synergic effect to elicit p53-, β-catenin- and MGMT-dependent apoptotic cell death.

doi: 10.1186/s12964-022-01007-x

Figure Lengend Snippet: Fig. 5 BMX, VPA, SAHA, TMZ, Oxp, and Dox combination inhibited cell proliferation in CRC cells. (A) HDAC8 mRNA expression levels stimulated with BMX (5 µM), VPA (4 mM), SAHA (2 µM) with or without TMZ were determined using qRT-PCR assays. (B) HT29, HCT116, and RKO cells were treated with BMX with or without TMZ for 48 h. Then, cells were harvested for detection of acetyl-histone H3 (Lys9/Lys14), acetyl-histone H4 (Lys8), and HDAC8 by Western blot analysis. (C) The proliferation of BMX, VPA, SAHA with or without TMZ, Oxp (5 µM) and Dox (1 µM) in HT29, HCT116, and RKO cells with treatment durations were assayed using the CCK-8 method. (D) Colony formation capability assay with different treatments of BMX, VPA, SAHA with or without TMZ, Oxp and Dox in HT29, HCT116, and RKO cells; the colonies were counted for quantification. All results are shown as mean ± s.d. from three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 versus control (HT29 cells); #p < 0.05, ##p < 0.01, ###p < 0.001 versus control (HCT116 cells); †p < 0.05, ††p < 0.01, †††p < 0.001 versus control (RKO cells)

Article Snippet: The cell proliferation of CRC cells was measured by CCK8 assay (Targetmol, Shanghai, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, CCK-8 Assay, Control

Journal: Cell communication and signaling : CCS

Article Title: BMX, a specific HDAC8 inhibitor, with TMZ for advanced CRC therapy: a novel synergic effect to elicit p53-, β-catenin- and MGMT-dependent apoptotic cell death.

doi: 10.1186/s12964-022-01007-x

Figure Lengend Snippet: Fig. 7 Autophagy expression levels stimulated with BMX and PCI-34051 in the presence or absence of TMZ. (A) HT29, HCT116, and RKO cells were treated with BMX and PCI-34051 with or without TMZ for 48 h. Then, cells were harvested for detection of acetyl-histone H3 (Lys9/Lys14), acetyl-histone H4 (Lys8), and HDAC8 by Western blot analysis. (B) The proliferation of BMX and PCI-34051 with or without TMZ in HT29, HCT116, and RKO cells. (C) Colony formation capability assay with different treatments of BMX and PCI-34051 with or without TMZ in HT29, HCT116, and RKO cells; the colonies were counted for quantification. (D, E) Expressions of cleaved caspase 3, cleaved PARP, LC3 and p62 proteins in HT29, HCT116, and RKO cells treated with BMX and PCI-34051 with or without TMZ for 48 h. All results are shown as mean ± s.d. from three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 versus control (HT29 cells); #p < 0.05, ##p < 0.01, ###p < 0.001 versus control (HCT116 cells); †p < 0.05, ††p < 0.01, †††p < 0.001 versus control (RKO cells)

Article Snippet: The cell proliferation of CRC cells was measured by CCK8 assay (Targetmol, Shanghai, China).

Techniques: Expressing, Western Blot, Control

Journal: Molecules

Article Title: Effects of Betaine on LPS-Stimulated Activation of Microglial M1/M2 Phenotypes by Suppressing TLR4/NF-κB Pathways in N9 Cells

doi: 10.3390/molecules24020367

Figure Lengend Snippet: Effects of betaine on LPS-induced TLR4/NF-κΒ signal transduction in N9 microglia cells. Cells were treated with betaine (1 mM) or MIDO (10 μM) for 1 h and then incubated with or without LPS (1 μg/mL) for 24 h. ( A ) Determination of TLR4/Myd88 protein levels by western blot analysis. ( B – D ) Determination of IKK-β, P-IKK-β, IκΒ-α, P-IκΒ-α, NF-κΒ, and p-NF-κΒ protein levels by western blot analysis. MIDO was used as a positive control. Data are presented as the means ± SEM of three independent experiments. The control group included untreated cells. Untreated cells served as a control group. # p < 0.05, compared to the control group; * p < 0.05 and ** p < 0.01, compared to the LPS-treated group.

Article Snippet: Cell extract (50 μg) was applied to a sodium dodecyl sulphate (SDS)-polyacrylamide gel electrophoresis, transferred to a polyvinylidene fluoride (PVDF) membrane, and then blocked with PBST containing 5% skimmed milk for 2 h. The membrane was incubated at 4 °C with primary antibodies (TLR4, 1:500; Myd88, 1:1000; iNOS, 1:1000; CD206, 1:2000; Arg-1, 1:1000 [Abcam]; IKK-β, 1:500; p-IKK-β, 1:200; IκBα, 1:1000; p-IκBα, 1:1000; NF-κB, 1:1000; p-NF-κB, 1:1000; GAPDH, 1:2000; α-tubulin, 1:1000; and β-tubulin, 1:1000 [Proteintech]) overnight.

Techniques: Transduction, Incubation, Western Blot, Positive Control

Journal: Oncotarget

Article Title: Identification of the histone lysine demethylase KDM4A/JMJD2A as a novel epigenetic target in M1 macrophage polarization induced by oxidized LDL

doi: 10.18632/oncotarget.17748

Figure Lengend Snippet: ( A ) RAW264.7 cells were exposed for 1 hr to 50 μg/ml oxLDL with or without pre-treatment with the JMJD inhibitor JIB-04 for 4 hrs, after which Western blot analysis was carried out to monitor expression of the indicated proteins. ( B ) RAW264.7 cells transfected with shKDM4A and scramble control shRNA were exposed to 50 μg/ml oxLDL for 1 hr, after which expression of HIF-1α, -1β, and phosphorylated p65 (S536) was monitored by Western blot analysis. ( C ) RAW264.7 cells were exposed for 1 hr to 50 μg/ml oxLDL with or without pre-treatment with the IKK inhibitor parthenolide (PTL) for 2 hrs, after which expression of phosphorylated p65 and KDM4A was assessed by Western blot analysis. ( D ) RAW264.7 cells were transiently transfected with pGreenpuro-shRNA constructs specifically targeting HIF-1β (shHIF-1β-1 and -2) or scrambled sequence as control. Down-regulation of HIF-1β was validated by Western blot analysis (upper panel). Cells were then exposed to 50 μg/ml oxLDL for 1 hr, after which expression of KDM4A was monitored by Western blot analysis.

Article Snippet: The IκB kinase (IKK) inhibitor parthenolide [ ] and the pan-selective Jumonji histone demethylase inhibitor JIB-04 [ ] were purchased from Merck Millipore (Billerica, MA).

Techniques: Western Blot, Expressing, Transfection, Control, shRNA, Construct, Sequencing