microplate based detection kit  (Elabscience Biotechnology)

Journal: International journal of molecular medicine

Article Title: Role of TXNDC5 in tumorigenesis of colorectal cancer cells: In vivo and in vitro evidence.

doi: 10.3892/ijmm.2018.3664

Figure Lengend Snippet: Figure 5. TXNDC5 promotes colorectal cancer cell proliferation, colony formation and survival in vitro. RKO and HCT‑116 cells were transfected with a specific siRNA (siTXNDC5) to knockdown TXNDC5. Cells transfected with siScr were used as a control. (A) MTT assays (*P<0.0001), (B and C) colony formation assays(*P<0.0001) and (D) apoptosis assays (**P=0.0020; ***P=0.0004) were performed in RKO and HCT‑116 cells transfected with siTXNDC5 or siScr under H and N. H/Hyp, hypoxia; N/Nom, normoxia; OD, optical density; siRNA/si, small interfering RNA; siScr, scrambled control siRNA; TXNDC5, thioredoxin domain‑containing 5.

Article Snippet: Cell apoptosis assay. cells were seeded at 1x105 cells/well in 96-well tissue culture plates. cell apoptosis was measured at 24 h using a microplate reader-based titertacS in situ apoptosis detection kit (r&d Systems, inc., minneapolis, mn, uSa) according to the manufacturer's protocol. the percentage of apoptotic cells was calculated.

Techniques: In Vitro, Transfection, Knockdown, Control, Small Interfering RNA

Journal: International journal of molecular medicine

Article Title: Role of TXNDC5 in tumorigenesis of colorectal cancer cells: In vivo and in vitro evidence.

doi: 10.3892/ijmm.2018.3664

Figure Lengend Snippet: Figure 7. TXNDC5 increases the expression of ER stress and apoptotic markers. (A) RKO and HCT‑116 cells were transfected with a specific siRNA (siTXNDC5) to knockdown TXNDC5. Cells transfected with siScr were used as a control. Reactive oxygen species production was mea- sured in cells under H and N for 24 h. *P=0.0004; **P=0.0010. (B) In RKO and HCT‑116 cells transfected with siTXNDC5 or siScr under H and N, western blot analyses were used to detect the protein expression levels of ER stress (CHOP, GRP78 and ATF4), apoptosis (Bax and cleaved caspase‑8) and survival (Bcl‑2) markers. β‑actin was used as a loading control. Density of the blots was normalized against that of the β‑actin blot, in order to obtain relative blot density. *P<0.05. ATF4, activating transcription factor 4; Bax, Bcl‑2‑associated X protein; Bcl‑2, B‑cell lymphoma 2; c‑C8, caspase‑8; CHOP, CCAAT‑enhancer‑binding protein homologous protein; GRP78, glucose‑regulated protein 78; DCF, dichlorodihydrofluorescein; ER, endoplasmic reticulum; H, hypoxia; N, normoxia; siRNA/si, small interfering RNA; siScr, scrambled control siRNA; TXNDC5, thioredoxin domain‑containing 5.

Article Snippet: Cell apoptosis assay. cells were seeded at 1x105 cells/well in 96-well tissue culture plates. cell apoptosis was measured at 24 h using a microplate reader-based titertacS in situ apoptosis detection kit (r&d Systems, inc., minneapolis, mn, uSa) according to the manufacturer's protocol. the percentage of apoptotic cells was calculated.

Techniques: Expressing, Transfection, Knockdown, Control, Western Blot, Small Interfering RNA