Journal: Asian Journal of Andrology

Article Title: Mannose inhibits the growth of prostate cancer through a mitochondrial mechanism

doi: 10.4103/aja2021104

Figure Lengend Snippet: Reagents applied in the study.

Article Snippet: Rhodamine 123 , MedChemExpress , Cat#HY-D8016.

Techniques: Enzyme-linked Immunosorbent Assay, ATP Assay, Transfection, Microarray

Journal: Asian Journal of Andrology

Article Title: Mannose inhibits the growth of prostate cancer through a mitochondrial mechanism

doi: 10.4103/aja2021104

Figure Lengend Snippet: Mannose disrupted mitochondrial function, led to ROS overproduction, and activated Bax/Bak in PCa cells. ( a ) JC-1 staining and ( b ) rhodamine 123 staining were used to assess the MMP in DU145 and PC3 cells. ( c ) The ATP content in cells. ( d ) Mitochondrial ROS and ( e ) cellular ROS levels in cells. ( f and g ) The protein expression of Bax and Bak in cells. * P < 0.05, ** P < 0.01. NC: PCa cells cultured in normal medium. MAN: PCa cells cultured in normal medium with 25 mmol l −1 mannose for DU145 or with 50 mmol l −1 mannose for PC3. JC-1: 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethyl-imidacarbocyanine; Bax: BCL2-associated X; Bak: BCL2-antagonist/killer 1; ROS: reactive oxygen species; MMP: mitochondrial membrane potential; ATP: adenosine triphosphate; PCa: prostate cancer.

Article Snippet: Rhodamine 123 , MedChemExpress , Cat#HY-D8016.

Techniques: Staining, Expressing, Cell Culture, Membrane

Journal: eLife

Article Title: Temporal transcription factors determine circuit membership by permanently altering motor neuron-to-muscle synaptic partnerships

doi: 10.7554/eLife.56898

Figure Lengend Snippet:

Article Snippet: 647-Phalloidin (1:600, Thermofisher A22287) Rhodamine-Phalloidin (1:600, Thermofisher R415), Cy3-HRP (Jackson ImmunoResearch 123-025-021), 647-HRP (Jackson ImmunoResearch 123-605-021).

Techniques: Western Blot

Journal: Cancer Science

Article Title: Identification of C2orf18, termed ANT2BP (ANT2‐binding protein), as one of the key molecules involved in pancreatic carcinogenesis

doi: 10.1111/j.1349-7006.2008.01058.x

Figure Lengend Snippet: C2orf18/ANT2BP was involved with mitochondrial membrane potential (Δψm) and apoptosis. (a) Pancreatic ductal adenocarcinoma (PDAC) cells KLM‐1 were transfected with adenine nucleotide translocase 2 (ANT2) small‐interfering RNA (siRNA), C2orf18/ANT2BP siRNA, or siEGFP (as a control) and collected 48 h after transfection. Western blot analysis using anti‐C2orf18/ANT2BP antibody confirmed the knockdown effect of C2orf18/ANT2BP siRNA on KLM‐1 cells. (b) The cells were incubated with Rhodamine123 (Rh123) and propidium iodide (PI), and fluorescence was measured by FACS analysis. Rh123 intensity on the x‐axis reflects Δψm, and PI permeability on the y‐axis reflects the cell membrane destruction in dead cells. The low level of Rh123 intensity and negative‐permeability of PI indicate the early apoptotic cells where Δψm is decreased but apoptosis does not finish completely, while the low level of Rh123 intensity and positive‐permeability of PI indicate dead cells. The numbers of the cells showing low Δψm and negative‐permeability of PI were increased when ANTBP2 (25%) or ANT2 (26%) were knocked down, compared with the control (siEGFP, 19%). (c) TUNEL assay showed that knockdown of C2orf18 in Panc‐1 cells increased the number of apoptosis cells (TUNEL‐positive cells indicated by green) compared with siEGFP‐transfected cells. The permeabilized cells treated with DNase I were prepared as positive controls for TUNEL assay. (d) The number of TUNEL‐positive cells was counted by flow cytometry. In the histogram plot, the x‐axis reflects the intensity of the green signals of TUNEL‐positive cells. The shift to the right of the histogram indicates that knockdown of C2orf18 increased the number of TUNEL‐positive cells compared with siEGFP‐transfected cells.

Article Snippet: The collected cells were washed twice with cold PBS, then incubated with 10‐µM Rhodamine123 (Rh123) (Wako, Osaka, Japan) in PBS for 15 min at 37°C in dark conditions, washed with fluorescence activated cell sorting (FACS) buffer, and resuspended in 0.5 mL of FACS buffer containing 10 µg/mL of propidium iodide (PI).

Techniques: Membrane, Transfection, Small Interfering RNA, Western Blot, Incubation, Fluorescence, Permeability, TUNEL Assay, Flow Cytometry