annexin v fitc pi kit  (Yeasen Biotechnology)

Journal: Oncology Reports

Article Title: Huaier promotes sensitivity of colorectal cancer to oxaliplatin by inhibiting METTL3 to regulate the Wnt/β‑catenin signaling pathway

doi: 10.3892/or.2024.8840

Figure Lengend Snippet: Knocking down METTL3 inhibits the Wnt/β-catenin signaling pathway, making colorectal cancer cells sensitive to OXA. (A) Western blot detection of METTL3 knockdown efficiency. (B) RT-qPCR detection of METTL3 knockdown efficiency. (C) Cell survival analysis of si-METTL3#1 and control si-NC treated with different concentrations of OXA. (D) Reverse transcription-quantitative PCR was used to detect the mRNA expression changes of P-gp and β-catenin after knocking down METTL3. (E) Flow cytometric analysis of cell apoptosis. (F) Western blot analysis was used to detect the expression changes of P-gp, Wnt 3a, β-catenin, Bax, Bcl-2 and cleaved caspase3/total caspase 3 proteins after knocking down METTL3. (G) Immunofluorescence analysis of β-catenin's entry into the nucleus. (H) Western blotting was used to detect the expression of β-catenin in the cytoplasm and nucleus of HCT-8/L cells. The data are expressed as the mean ± SD. *P<0.05, **P<0.01 and ***P<0.001 compared with the si-NC group. METTL3, methyltransferase 3; OXA, oxaliplatin; si-, small interfering; NC, negative control.

Article Snippet: After centrifugation at 1,000 × g for 5 min at room temperature, the supernatant was discarded completely and staining was performed with the Annexin V-FITC/PI dual staining apoptosis detection kit (APeXBIO Technology LLC), avoiding light at room temperature until 5 min.

Techniques: Western Blot, Knockdown, Quantitative RT-PCR, Control, Reverse Transcription, Real-time Polymerase Chain Reaction, Expressing, Immunofluorescence, Negative Control

Journal: Oncology Reports

Article Title: Huaier promotes sensitivity of colorectal cancer to oxaliplatin by inhibiting METTL3 to regulate the Wnt/β‑catenin signaling pathway

doi: 10.3892/or.2024.8840

Figure Lengend Snippet: Knocking down METTL3 increases the sensitivity of HCT-8/L cells to oxaliplatin by inhibiting the Wnt/β-catenin signaling pathway. (A) Flow cytometric analysis of cell apoptosis. (B) Reverse transcription-quantitative PCR was used to detect changes in mRNA expression of METTL3, P-gp and β-catenin. (C) Western blotting was used to detect the protein expression changes of METTL3, P-gp, Wnt 3a, β-catenin, Bax, Bcl-2 and cleaved caspase3/total caspase 3. (D) Immunofluorescence analysis of β-catenin's entry into the nucleus. (E) Western blot analysis of the expression of β-catenin in the cytoplasm and nucleus of HCT-8/L cells. The data are expressed as the mean ± SD. *P<0.05, **P<0.01 and ***P<0.001 compared with the si-NC group; # P<0.05, ## P<0.01 and #### P<0.0001 compared with the si-METTL3#1 group. METTL3, methyltransferase 3; si-, small interfering; NC, negative control; ns, not significant.

Article Snippet: After centrifugation at 1,000 × g for 5 min at room temperature, the supernatant was discarded completely and staining was performed with the Annexin V-FITC/PI dual staining apoptosis detection kit (APeXBIO Technology LLC), avoiding light at room temperature until 5 min.

Techniques: Reverse Transcription, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Immunofluorescence, Negative Control

Journal: Oncology Reports

Article Title: Huaier promotes sensitivity of colorectal cancer to oxaliplatin by inhibiting METTL3 to regulate the Wnt/β‑catenin signaling pathway

doi: 10.3892/or.2024.8840

Figure Lengend Snippet: Huaier downregulates the expression of METTL3, inhibits the Wnt/β-catenin signaling pathway, and renders HCT-8/L cells sensitive to OXA. (A) Survival analysis of HCT-8/L cells treated with different concentrations of OXA after treatment with Huaier. (B) Reverse transcription-quantitative PCR was used to detect the effect of Huaier on the mRNA expression of METTL3, P-gp and β-catenin. (C) Flow cytometric analysis of cell apoptosis. (D) Western blot was used to detect the effects of Huaier on the protein expression of METTL3, P-gp, Wnt 3a, β-catenin, Bax, Bcl-2 and cleaved caspase3/total caspase 3. (E) Immunofluorescence analysis of β-catenin's entry into the nucleus. (F) Western blotting was used to detect the expression of β-catenin in the cytoplasm and nucleus of HCT-8/L cells. The data are expressed as the mean ± SD. *P<0.05, **P<0.01 and ***P<0.001 compared with the HCT-8/L group. METTL3, methyltransferase 3; OXA, oxaliplatin.

Article Snippet: After centrifugation at 1,000 × g for 5 min at room temperature, the supernatant was discarded completely and staining was performed with the Annexin V-FITC/PI dual staining apoptosis detection kit (APeXBIO Technology LLC), avoiding light at room temperature until 5 min.

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Immunofluorescence

Journal: Oncology Reports

Article Title: Huaier promotes sensitivity of colorectal cancer to oxaliplatin by inhibiting METTL3 to regulate the Wnt/β‑catenin signaling pathway

doi: 10.3892/or.2024.8840

Figure Lengend Snippet: Huaier inhibits the Wnt/β-catenin signaling pathway by downregulating the expression of METTL3, rendering HCT-8/L cells sensitive to OXA. (A) Western blotting was used to detect the overexpression efficiency of METTL3. (B) RT-qPCR was used to detect the overexpression efficiency of METTL3. (C) Flow cytometric analysis of cell apoptosis. (D) RT-qPCR was used to detect changes in mRNA expression of METTL3, P-gp and β-catenin. (E) Western blotting was used to detect the protein expression changes of METTL3, P-gp, Wnt 3a, β-catenin, Bax, Bcl-2 and cleaved caspase3/total caspase 3. (F) Immunofluorescence analysis of β-catenin's entry into the nucleus. (G) Western blot analysis of the expression of β-catenin in the cytoplasm and nucleus of HCT-8/L cells. The data are expressed as the mean ± SD. *P<0.05, **P<0.01, ***P<0.001 and ****P<0.0001 compared with the pC-NC group; # P<0.05, ## P<0.01, ### P<0.001 and #### P<0.0001 compared with the pC-METTL3. METTL3, methyltransferase 3; OXA, oxaliplatin; RT-qPCR, reverse transcription-quantitative PCR; NC, negative control; ns, not significant.

Article Snippet: After centrifugation at 1,000 × g for 5 min at room temperature, the supernatant was discarded completely and staining was performed with the Annexin V-FITC/PI dual staining apoptosis detection kit (APeXBIO Technology LLC), avoiding light at room temperature until 5 min.

Techniques: Expressing, Western Blot, Over Expression, Quantitative RT-PCR, Immunofluorescence, Reverse Transcription, Real-time Polymerase Chain Reaction, Negative Control

Journal: Oncology Letters

Article Title: Potential antitumor effect of polysaccharides extracted from Polygonatum sibiricum on human prostate cancer PC‑3 cells

doi: 10.3892/ol.2024.14774

Figure Lengend Snippet: PSP promotes cell apoptosis and caspase-3 expression. PC-3 cells were treated with different concentrations of PSP for 72 h. (A) Cell apoptosis analyzed using flow cytometry, with apoptotic cells distributed in the Q1-2 and Q1-4 quadrants of the charts. (B) Apoptotic rates in the PSP-treated groups, representing the sum of Q1-2 and Q1-4, which were significantly higher than in the blank control group. (C) Lysates from treated cells were collected and the expression of apoptosis-related protein caspase-3 was detected using western blotting. (D) Semi-quantitative analysis of caspase-3 revealed a concentration-dependent increase in caspase-3 activity. *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001. PSP, Polygonatum sibiricum .

Article Snippet: Based on the results of the CCK-8 assay, PC-3 cells were plated into 6-well plates at a density of 2×10 5 cells/well and treated with several concentrations of PSP solution (0, 250, 500 µg/ml, 1, 2 and 4 mg/ml) at 37°C for 72 h. The apoptosis rate of PC-3 cells was evaluated using the Annexin V-FITC/PI Apoptosis Detection Kit according to the manufacturer's instructions (Beijing 4A Biotech Co., Ltd. Cat. FXP018).

Techniques: Expressing, Flow Cytometry, Control, Western Blot, Concentration Assay, Activity Assay