Journal: Cells

Article Title: HDAC1 -Mediated lncRNA Stimulatory Factor of Follicular Development to Inhibit the Apoptosis of Granulosa Cells and Regulate Sexual Maturity through miR-202-3p- COX1 Axis

doi: 10.3390/cells12232734

Figure Lengend Snippet: HDAC1 regulates the proliferation and apoptosis of GCs. ( A ) Effects TSA treatment at concentration of 0.01, 0.1, 0.25, 0.5, and 1 μM for 24 and 48 h on the viability of porcine GCs. ( B ) Micrographs of GCs treated with 0.1 and 0.25 μM TSA at 24 and 48 h. ( C ) Effects of 0.01, 0.1, 0.25, 0.5, and 1 μM TSA treatment on the expression of HDAC1 for 24 and 48 h. ( D – F ) The mRNA and protein levels of cell cycle and cell apoptosis-related genes in GCs treated with TSA. ( G , H ) The proliferation and apoptosis of GCs treated with TSA were assessed using EdU and flow cytometry. ( I ) Effect of TSA on the activity of Caspase 3/7 in porcine GCs was detected using Caspase 3/7 activity assay. ( J ) The interference efficiency of HDAC1 . ( K – M ) The mRNA and protein levels of cell cycle and cell apoptosis-related genes in GCs treated with HDAC1 -siRNA. ( N , O ) The proliferation and apoptosis of GCs treated with HDAC1 -siRNA were assessed using EdU and flow cytometry. ( P ) Effect of HDAC1 -siRNA on the activity of Caspase 3/7 in porcine GCs was detected using Caspase 3/7 activity assay. * indicates p < 0.05, ** indicates p < 0.01.

Article Snippet: The Caspase 3/7 Activity Assay was also performed to detect the apoptosis of GCs using the Caspase 3/7 Activity Assay Kit (Elabscience, Wuhan, China).

Techniques: Concentration Assay, Expressing, Flow Cytometry, Activity Assay

Journal: Cells

Article Title: HDAC1 -Mediated lncRNA Stimulatory Factor of Follicular Development to Inhibit the Apoptosis of Granulosa Cells and Regulate Sexual Maturity through miR-202-3p- COX1 Axis

doi: 10.3390/cells12232734

Figure Lengend Snippet: SFFD as ceRNA for miR-202-3p- COX1 regulates the proliferation and apoptosis of porcine GCs. ( A – D ) Effects of SFFD overexpression and miR-202-3p mimic on the mRNA and protein levels of genes related to cell cycle, E2 secretion, and apoptosis. ( E – H ) Effects of SFFD knockdown and miR-202-3p inhibitor on the mRNA and protein levels of genes related to cell cycle, E2 secretion, and apoptosis. ( I ) Effects of SFFD overexpression and miR-202-3p mimic on the proliferation of GCs. ( J ) Effects of SFFD knockdown and miR-202-3p inhibitor on the proliferation of GCs. ( K ) Effects of SFFD overexpression with miR-202-3p mimic and SFFD knockdown with miR-202-3p inhibitor on the E2 secretion of GCs. ( L , M ) Effects of SFFD overexpression and miR-202-3p mimic on the apoptosis and Caspase 3/7 activity of GCs. ( N , O ) Effects of SFFD knockdown and miR-202-3p inhibitor on the apoptosis and Caspase 3/7 activity of GCs. * indicates p < 0.05, ** indicates p < 0.01.

Article Snippet: The Caspase 3/7 Activity Assay was also performed to detect the apoptosis of GCs using the Caspase 3/7 Activity Assay Kit (Elabscience, Wuhan, China).

Techniques: Over Expression, Knockdown, Activity Assay

Journal: Cancer Science

Article Title: Peneciraistin C induces caspase‐independent autophagic cell death through mitochondrial‐derived reactive oxygen species production in lung cancer cells

doi: 10.1111/cas.12253

Figure Lengend Snippet: Peneciraistin C (Pe‐C) induces caspase‐independent non‐apoptotic cell death in three lung cancer cell lines. (a) A549, H446, H661, and BEAS‐2B cells were treated with indicated concentrations of Pe‐C for 24 and 48 h. Cell viabilities were determined by MTT assay. The IC 50 value was calculated by an IC 50 software program. (b) A549, H446, and H661 cells were treated with indicated concentrations of Pe‐C and 5‐fluorouracil (5‐FU) in the absence (–) or presence of pan‐caspase inhibitor z‐VAD‐fmk (100 μM) for 48 h. The percentage of cell death was quantified by Trypan blue staining. (c) A549 cells were treated with indicated concentrations of Pe‐C or 5‐FU for the indicated times. The activity of caspase‐3, ‐8, and ‐9 was measured by a microplate reader at 405 nm. (d) A549 cells were treated with indicated concentrations of Pe‐C or 5‐FU for 24 and 48 h. Cells were stained with propidium iodide and analyzed by flow cytometry. Data represent results of at least three independent experiments.

Article Snippet: The activity of caspase‐3, ‐8, and ‐9 was measured using commercially available caspase colorimetric assay kits (Beyotime).

Techniques: MTT Assay, Software, Staining, Activity Assay, Flow Cytometry

Journal: Cancer Science

Article Title: Peneciraistin C induces caspase‐independent autophagic cell death through mitochondrial‐derived reactive oxygen species production in lung cancer cells

doi: 10.1111/cas.12253

Figure Lengend Snippet: Blockage of autophagy converts peneciraistin C (Pe‐C)‐induced cell death to apoptotic cell death. A549 cells were pretreated with 3‐methyladenine (3‐MA, 5 mM) or wortmannin (WM, 100 nM) for 1 h, followed by 3 μM Pe‐C for the indicated times. (a) After treatment for 48 h, whole cell lysates were separated by SDS‐PAGE, then immunoblotting was carried out using anti‐LC3 antibody. (b) Cell viabilities were analyzed by MTT assay. (c) Sub‐G1 cell cycle phase was analyzed using flow cytometry. (d) Caspase‐3 activity was measured by a microplate reader at 405 nm.

Article Snippet: The activity of caspase‐3, ‐8, and ‐9 was measured using commercially available caspase colorimetric assay kits (Beyotime).

Techniques: SDS Page, Western Blot, MTT Assay, Flow Cytometry, Activity Assay