Journal: Molecules

Article Title: Inhibition of MORC2 Mediates HDAC4 to Promote Cellular Senescence through p53/p21 Signaling Axis

doi: 10.3390/molecules27196247

Figure Lengend Snippet: Knockdown of MORC2 inhibits HCT-116 and Lovo cell proliferation and clone formation. ( A ) GSEA indicates that MORC2-associated DEGs were enriched in the cell cycle. ( B ) HCT-116 and Lovo cells were transfected with indicated shRNA, and the protein expression was analyzed with indicated antibodies. ( C ) CCK8 assays were used to assess the DNA synthesis of shCON, shMORC2#1, and shMORC2#2 groups with HCT-116 and Lovo cells. ( D , E ) Edu assay was used to assess the DNA synthesis of shCON, shMORC2#1, and shMORC2#2 groups with HCT-116 cells. The percentage of EdU-positive cells in shCON, shMORC2#1, and shMORC2#2 groups. ( F ) Colony formation assay showing clone formation capacity of shCON, shMORC2#1, and shMORC2#2 groups. ( G ) Each group colony number was counted and relative colony numbers in shMORC2#1 and shMORC2#2 were compared to the shCON group. ** p < 0.01, **** p < 0.0001.

Article Snippet: HCT-116 cells were obtained from Procell Co., Ltd. HCT-116 and HEK293 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS), under 37 °C and 5% CO 2 conditions.

Techniques: Transfection, shRNA, Expressing, DNA Synthesis, EdU Assay, Colony Assay

Journal: Molecules

Article Title: Inhibition of MORC2 Mediates HDAC4 to Promote Cellular Senescence through p53/p21 Signaling Axis

doi: 10.3390/molecules27196247

Figure Lengend Snippet: Knockdown of MORC2 induces cell senescence. ( A ) PPI network of the top 10 interacting genes related to MORC2. ( B ) GSEA indicates that MORC2-associated DEGs were enriched in cell senescence. ( C ) The single gene co-expression heat map presented the significant relationship among MORC2, HDAC4, IL-1α, and IL-1β. ( D ) HCT-116 and Lovo cells are transfected with shMORC2, and the protein expression is analyzed with indicated antibodies. ( E ) p53 promoter-luciferase activity was measured. ( F ) mRNA expression of p16, IL-1α, and IL-1β in HCT-116 cells. ( G – J ) The levels of IL-6 and IL-8 secreted by HCT-116 and Lovo cells after transfecting with shMORC2. ( K ) SA-β-gal staining analysis in HCT-116 and Lovo cells transfected with shMORC2. ( L ) Relative fold change of the SA-β-gal staining ratio in each shMORC2 group was calculated and compared to shCON group’s SA-β-gal staining ratio. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: HCT-116 cells were obtained from Procell Co., Ltd. HCT-116 and HEK293 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS), under 37 °C and 5% CO 2 conditions.

Techniques: Expressing, Transfection, Luciferase, Activity Assay, Staining

Journal: Molecules

Article Title: Inhibition of MORC2 Mediates HDAC4 to Promote Cellular Senescence through p53/p21 Signaling Axis

doi: 10.3390/molecules27196247

Figure Lengend Snippet: Knockdown of MORC2 inhibits tumor growth. ( A ) The growth curve of mice xenografts formed by shCON and shMORC2#1 cells ( n = 5). ( B ) Tumor weights after sacrificing the mice ( n = 5). ( C ) Mice xenografts formed by HCT-116 with or without MORC2 intervention. ( D , E ) IHC staining and statistical analysis of xenografts using antibodies against ki67, MORC2, p53, and HDAC4. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: HCT-116 cells were obtained from Procell Co., Ltd. HCT-116 and HEK293 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS), under 37 °C and 5% CO 2 conditions.

Techniques: Immunohistochemistry

Journal: The Journal of Biological Chemistry

Article Title: Calcium Co-regulates Oxidative Metabolism and ATP Synthase-dependent Respiration in Pancreatic Beta Cells

doi: 10.1074/jbc.M113.513184

Figure Lengend Snippet: Glucose- and calcium-mediated activation of ATP synthase-dependent respiration in human islets. Human islets bound to 804G matrix were incubated in KRBH ( A ) or a KRBH buffer lacking Ca 2+ supplemented with 0.4 m m EGTA ( B ; Ca 2+ -free). For each condition the mean ± S.E. n = 3 from the same donor is shown. Total islet protein varied between wells (4–6 μg). Because of these variations the results are expressed relative to the respiratory rate before glucose stimulation. ATP synthase-dependent ( dep , C ) and -independent respiration ( D ) was quantified as described under “Experimental Procedures.” The results are the mean ± S.E. ( n = 6) obtained from 2 donors. *, p < 0.01; **, p < 0.001; ns , not significant. R , rotenone (1 μ m ); AA , antimycin A (1 μg/ml); O , oligomycin (2.5 μg/ml); glc , glucose. E , preventing calcium signaling blocks glucose-induced insulin secretion. Insulin secretion from human islets was determined in KRBH or the same buffer lacking Ca 2+ in either 1 m m ( gray bars ) or 16.7 m m glucose ( black bars ). Shown is the average ±S.E. n = 4 result with islets from a single donor (*, p < 0.05).

Article Snippet: The 804G bladder carcinoma cell line ( ) and protocols to isolate the matrix were obtained from Viacyte (San Diego, CA).

Techniques: Activation Assay, Incubation