Journal: Journal of Clinical Medicine

Article Title: Diets Differently Regulate Tumorigenesis in Young E0771 Syngeneic Breast Cancer Mouse Model

doi: 10.3390/jcm12020413

Figure Lengend Snippet: HFD increases autophagic apoptosis at the early tumor point and decreases it at a later stage of tumor growth as demonstrated by IHC in tumor sections. ( A ) The markers of autophagy (LC3) and apoptosis (Annexin V) significantly increased in short-term HFD-fed mice compared to RD-fed mice at 2 wpi; ( B ) however, the levels of these markers significantly decreased at 5 wpi in both RD- and HFD-fed mice compared to 2 wpi (black arrows indicate the invasive front associated with adipocytes) (40× magnification).

Article Snippet: Immunohistochemistry (IHC) was performed on tumor sections using rabbit polyclonal LC3A/LC3B antibody (#PA1-16931, Invitrogen, Waltham, MA, USA) and rabbit polyclonal Annexin V antibody (#8555, Cell Signaling Technology, Danvers, MA, USA) with a dilution of 1:200 and 1:300, respectively, followed by biotinylated secondary antibody using VECTASTAIN Elite ABC-HRP kit (#PK-6101, Vector Laboratories, Burlingame, CA, USA).

Techniques:

Journal: Materials Today Bio

Article Title: Bone tumor-homing nanotherapeutics for prolonged retention in tumor microenvironment and facilitated apoptotic process via mevalonate pathway inhibition

doi: 10.1016/j.mtbio.2023.100591

Figure Lengend Snippet: Analyses of the apoptosis and mevalonate pathway inhibition in PLCSA-AD-treated HOS/MNNG cells. (a) Representative flow cytometry contour plots of HOS/MNNG cells stained with annexin V-FITC and PI. (b) Western blot analysis of representative apoptosis-related proteins after treatment with NTs. (c, d) Western blot analyses of (c) small GTPases and (d) the phosphorylated ERK1/2 and MEK after treatment with blank PLCSA or PLCSA-AD. The quantification of the Western blot images is available in the Supplementary Materials as Fig. S4 .

Article Snippet: The cells and supernatant were collected and stained with the Annexin V-FITC Early Apoptosis Detection Kit (Cell Signaling Technology) according to the manufacturer's protocol.

Techniques: Inhibition, Flow Cytometry, Staining, Western Blot

Journal: Foods

Article Title: In vitro Digestion of Phaseolus vulgaris L. Cooked Beans Induces Autophagy in Colon Cancer Cells

doi: 10.3390/foods12040839

Figure Lengend Snippet: BF does not induce colon cancer cell death through the activation of apoptotic process. HT29 cells were incubated with 100 µg/mL of Cannellino and Piattellino BF and stained with ( A ) 0.1 µg/mL DAPI or ( B ) Annexin V-FITC conjugated antibody and Propidium Iodide. Data are representative of three independent experiments.

Article Snippet: After 24 h, cells were treated with Cannellino or Piattellino BF (100 µg/mL) for 18 h. Each well was washed with PBS and incubated with 1X Annexin V Binding Buffer containing 1 µL of Annexin V-FITC conjugated antibody and 12.5 µL of propidium iodide (cell signaling) for 10 min on ice in the dark.

Techniques: Activation Assay, Incubation, Staining

Journal: Foods

Article Title: In vitro Digestion of Phaseolus vulgaris L. Cooked Beans Induces Autophagy in Colon Cancer Cells

doi: 10.3390/foods12040839

Figure Lengend Snippet: BF does not induce colon cancer cell death through the activation of apoptotic process. HT29 cells were incubated with 100 µg/mL of Cannellino and Piattellino BF and stained with ( A ) 0.1 µg/mL DAPI or ( B ) Annexin V-FITC conjugated antibody and Propidium Iodide. Data are representative of three independent experiments.

Article Snippet: After 24 h, cells were treated with Cannellino or Piattellino BF (100 µg/mL) for 18 h. Each well was washed with PBS and incubated with 1X Annexin V Binding Buffer containing 1 µL of Annexin V-FITC conjugated antibody and 12.5 µL of propidium iodide (cell signaling) for 10 min on ice in the dark.

Techniques: Activation Assay, Incubation, Staining

Journal: Biomedicines

Article Title: Targeting Periostin Expression Makes Pancreatic Cancer Spheroids More Vulnerable to Natural Killer Cells

doi: 10.3390/biomedicines11020270

Figure Lengend Snippet: Cell death was detected to increase in periostin-suppressed spheroids. ( A ) Annexin-V and PI positivity in total cell populations. The graphs show the percentage of Annexin-V and PI positivity in BxPC-3 cells mono-culture, BxPC-3+PSC co-culture and BxPC-3+PSC-P co-culture spheroids following 24 h exposure to lenalidomide-stimulated NK-92 cells. ( B ) Annexin-V and PI positivity in EpCAM(+) cancer cell population. The graphs show the percentage of Annexin-V and PI staining in EpCAM(+) BxPC-3 mono-culture, BxPC-3+PSC, and BxPC-3+PSC-P co-culture spheroids were exposed to lenalidomide-stimulated NK-92 cells for 24 h.

Article Snippet: Cells were kept in the dark at RT for 20 min. Live/Dead cell analysis, Annexin-V-FITC (1:100, Cell Signaling Technology, Danvers, MA, USA) and Propidium Iodide-PE (1:10, Cell Signaling Technology, USA) were used.

Techniques: Co-Culture Assay, Staining

Journal: Life Science Alliance

Article Title: Trans-differentiation of trophoblast stem cells: implications in placental biology

doi: 10.26508/lsa.202201583

Figure Lengend Snippet: (A, B, C) Quantitative real-time PCR (qRT) analysis of the differentially expressed endothelial cell–specific genes in trophoblast stem cells (TS) and differentiated trophoblast cells (TC). qRT–PCR results are grouped based on functional annotations. (A) Angiogenesis-related genes: Cx3cl1,c-kit , Kdr , Plau , Mmp . (B) Cell adhesion molecules: Cdh5 , Pecam1 , Itg β3, Col18a1 . (C) Apoptosis-related genes: Tnfsf10 , Bcl2 , Cradd , Casp3 . Data are representative of three biological replicates, and error bars represent SEM. * P < 0.5, ** P < 0.01. Source data are available for this figure.

Article Snippet: After 48 and 72 h, co-cultured cells were processed either for both annexin V–PI staining using early apoptosis detection kit (cat no. 6592; Cell Signaling Technology) by flow cytometry or for protein isolation and Western blotting for detection of apoptosis markers.

Techniques: Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Functional Assay

Journal: Life Science Alliance

Article Title: Trans-differentiation of trophoblast stem cells: implications in placental biology

doi: 10.26508/lsa.202201583

Figure Lengend Snippet: (A) Apoptotic death induced by increasing dose of TRAIL in endothelial cells (MS1) and differentiated trophoblast cells (TC) assessed using annexin V–PI–based flow cytometric analysis. Corresponding untreated cells were kept as controls. (A, B, C) Quantitative analysis of percentage of from (A) in MS1 cells and (C) TC has been shown in bar graphs. Data are representative of three independent biological replicates. Error bar represents SEM. *** P < 0.001; ns, nonsignificant. (D, E) Annexin V–PI–based flow cytometric analysis to assess apoptotic death of endothelial cells (MS1) co-cultured either in the absence or presence of differentiated trophoblast cells for 48 h (D) or 72 h (E). Quantitative analysis of percentage of cells undergoing apoptotic death has been shown in adjacent bar graphs. (F) Western blot analysis of the TNFSF10 (TRAIL) agonistic receptor DR4 using cell lysate from MS1 co-cultured either in the absence (control) or presence of differentiated trophoblast cells and cell lysate from co-cultured trophoblast cells (TC). (F, G) Densitometric analysis of the proteins from (F) using NIH ImageJ software after normalization with GAPDH. Data are representative of three independent biological replicates. (H, J) Western blot analysis of proteins from extrinsic (H) and intrinsic (J) apoptotic pathway proteins using lysates from MS1 cells co-cultured in the absence or presence of TC. (G, I, K) Densitometric analysis of the proteins from (G, I), respectively, using NIH ImageJ software after normalization with GAPDH. Data are representative of three independent biological replicates. Error bars represent SEM. ** P < 0.01; ns, nonsignificant. Source data are available for this figure.

Article Snippet: After 48 and 72 h, co-cultured cells were processed either for both annexin V–PI staining using early apoptosis detection kit (cat no. 6592; Cell Signaling Technology) by flow cytometry or for protein isolation and Western blotting for detection of apoptosis markers.

Techniques: Cell Culture, Western Blot, Software

Journal: Life Science Alliance

Article Title: Trans-differentiation of trophoblast stem cells: implications in placental biology

doi: 10.26508/lsa.202201583

Figure Lengend Snippet: (A) Apoptotic death induced by increasing dose of TRAIL in endothelial cells (MS1) and differentiated trophoblast cells (TC) assessed using annexin V–PI–based flow cytometric analysis. Corresponding untreated cells were kept as controls. (A, B, C) Quantitative analysis of percentage of from (A) in MS1 cells and (C) TC has been shown in bar graphs. Data are representative of three independent biological replicates. Error bar represents SEM. *** P < 0.001; ns, nonsignificant. (D, E) Annexin V–PI–based flow cytometric analysis to assess apoptotic death of endothelial cells (MS1) co-cultured either in the absence or presence of differentiated trophoblast cells for 48 h (D) or 72 h (E). Quantitative analysis of percentage of cells undergoing apoptotic death has been shown in adjacent bar graphs. (F) Western blot analysis of the TNFSF10 (TRAIL) agonistic receptor DR4 using cell lysate from MS1 co-cultured either in the absence (control) or presence of differentiated trophoblast cells and cell lysate from co-cultured trophoblast cells (TC). (F, G) Densitometric analysis of the proteins from (F) using NIH ImageJ software after normalization with GAPDH. Data are representative of three independent biological replicates. (H, J) Western blot analysis of proteins from extrinsic (H) and intrinsic (J) apoptotic pathway proteins using lysates from MS1 cells co-cultured in the absence or presence of TC. (G, I, K) Densitometric analysis of the proteins from (G, I), respectively, using NIH ImageJ software after normalization with GAPDH. Data are representative of three independent biological replicates. Error bars represent SEM. ** P < 0.01; ns, nonsignificant. Source data are available for this figure.

Article Snippet: Cells were then stained with annexin V and PI (cat no. 6592; Cell Signaling Technology) as per manufacturer’s instructions.

Techniques: Cell Culture, Western Blot, Software