Journal: Oncology Research

Article Title: RNA Interference of Biot2 Induces G 1 Phase Arrest and Apoptosis in Mouse Colorectal Cancer Cell Line

doi: 10.3727/096504014X14146137738583

Figure Lengend Snippet: Biot2 genes downregulated arrest of CT26 cell cycle. The effect of Biot2-shRNA on inducing G 1 cell cycle arrest in CT26 cells. After treatment with Biot2-shRNA for 48 h, (A) the cell cycle distributions were determined by flow cytometry. (B) The protein expressions of cycle regulatory proteins and cyclin-dependent kinase cyclin D1, CDK2 and (C) the expression of several CKIs, p16, p21, and p27, were measured by Western blotting.

Article Snippet: Rabbit anti-CDK2 (1:1,000), rabbit anti-p16 (1:1,000), rabbit anti-p27 (1:1,000), and mouse anti-PCNA (1:1,000) were purchased from BOSTER (China).

Techniques: shRNA, Flow Cytometry, Expressing, Western Blot

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Retroviral host range extension is coupled with Env-activating mutations resulting in receptor-independent entry

doi: 10.1073/pnas.1704750114

Figure Lengend Snippet: Virus harboring EnvH20 has a lower titer than the virus with original EnvC despite the same RT activity and VLP formation. (A) Infection of NIL-Tvc and DF-1 cells with RCAS-EnvC-GFP and RCAS-EnvH20-GFP viruses was scored by flow cytometry 2 (DF-1 cell) or 3 (NIL-Tvc) days later. Titers were determined as described in Materials and Methods. The results are presented as titers relative to RCAS-EnvC-GFP. (B) RT activity of RCAS-EnvC and RCAS-EnvH20-GFP was measured using the PERT assay described in SI Materials and Methods. The results of the PERT assay are presented relative to RCAS-EnvC-GFP. (C) VLP production of DF-1 cells infected with RCAS-EnvC-GFP or RCAS-EnvH20-GFP was estimated by Western blot analysis. Expression of Gag product p27 in the medium was determined with anti-p27 antibody.

Article Snippet: The presence of virions was detected by immunoblotting with the anti-ASLV rabbit p27 Antiserum (Charles River Laboratories, 1:1,000).

Techniques: Virus, Activity Assay, Infection, Flow Cytometry, Western Blot, Expressing

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Retroviral host range extension is coupled with Env-activating mutations resulting in receptor-independent entry

doi: 10.1073/pnas.1704750114

Figure Lengend Snippet: EnvH20 has similar features as a receptor-primed Env. (A) The virus with EnvH20 is thermosensitive. Virus stocks of similar titers were incubated at 44 °C for the indicated time. The titers were determined on NIL-Tvc 3 d after infection using flow cytometry. The amount of proviral DNA in infected DF-1 cells was measured 1 d after infection using RT-PCR. The results are presented as the percentage of the original titer (full lines) or the amount of DNA (dashed lines) remaining after heating. (B) The virus with EnvH20 is inactivated by low pH. Purified viruses of similar titers were incubated with low (pH 5) or neutral (pH 7.5) treatment at 37 °C for 30 min before infection of DF-1 cells. The titers were determined by flow cytometry 2 d later. (C) Formation of TM oligomers triggered by increasing temperature. The virus was incubated for 20 min at the indicated temperature. Samples were lysed in Laemmli loading buffer and analyzed by SDS/PAGE without boiling. TM was detected by immunoblotting with an antibody against its C-terminal part. Only the 70-kDa isoform of TM is shown. (D) Formation of TM oligomers at low pH. The virus was incubated for 30 min at the indicated pH at room temperature. Samples were neutralized, lysed in Laemmli loading buffer, and analyzed by SDS/PAGE without boiling. TM was detected by immunoblotting with an antibody against its C-terminal part. (E) The virus with EnvH20 binds liposomes. Viruses of similar titers were incubated with or without liposomes at 37 °C and then the mixture was separated by sucrose gradient centrifugation. Sample fractions were collected and analyzed by Western immunoblotting using anti-p27 antibody. T, top; B, bottom. (F) The virus containing EnvH20 is inhibited with PMB. Purified viruses of similar titers were incubated at 37 °C for 30 min with increasing concentration of PMB. Viruses were diluted in medium and spinoculated on DF-1 cells. The percentage of GFP+ cells was measured by flow cytometry 2 d after infection. Titers below the limit of detection are marked with “<”. Error bars show the SD of the two independent experiments in parallel. Significant differences are marked by asterisks (*P = 0.05–0.01, **P = 0.01–0.001, ***P < 0.001). NS, not significant.

Article Snippet: The presence of virions was detected by immunoblotting with the anti-ASLV rabbit p27 Antiserum (Charles River Laboratories, 1:1,000).

Techniques: Virus, Incubation, Infection, Flow Cytometry, Reverse Transcription Polymerase Chain Reaction, Purification, SDS Page, Western Blot, Liposomes, Gradient Centrifugation, Concentration Assay

Journal: Endocrine Connections

Article Title: The emerging role of the molecular marker p27 in the differential diagnosis of adrenocortical tumors

doi: 10.1530/EC-13-0025

Figure Lengend Snippet: Percentage area stained for the different immunohistochemical markers (mean± s.e.m .) in the different groups.

Article Snippet: The sections were incubated overnight at 4 °C with the appropriate diluted primary antibodies: rabbit anti-human MABs to p53 (453M-94; 1:100; Cell Marque, Rocklin, CA, USA), p21 (421M-14; 1:50; Cell Marque), p27 (427M-94; 1:500; Cell Marque), cyclin D1 (271R-14; 1:500; Cell Marque), Ki-67 (27R-14; 1:100; Cell Marque), and E-cadherin (246R-14; 1:200; Cell Marque) and rabbit anti-human polyclonal antibodies to β-catenin (424A-14; 1:500; Cell Marque).

Techniques: Staining, Immunohistochemical staining

Journal: Endocrine Connections

Article Title: The emerging role of the molecular marker p27 in the differential diagnosis of adrenocortical tumors

doi: 10.1530/EC-13-0025

Figure Lengend Snippet: Graphical representation of ROC curves with the respective area under the curve (AUC) to compare carcinoma (ACC) and adenoma with Cushing's syndrome (ACAc) samples for the markers StAR, p27, and Ki-67.

Article Snippet: The sections were incubated overnight at 4 °C with the appropriate diluted primary antibodies: rabbit anti-human MABs to p53 (453M-94; 1:100; Cell Marque, Rocklin, CA, USA), p21 (421M-14; 1:50; Cell Marque), p27 (427M-94; 1:500; Cell Marque), cyclin D1 (271R-14; 1:500; Cell Marque), Ki-67 (27R-14; 1:100; Cell Marque), and E-cadherin (246R-14; 1:200; Cell Marque) and rabbit anti-human polyclonal antibodies to β-catenin (424A-14; 1:500; Cell Marque).

Techniques:

Journal: Endocrine Connections

Article Title: The emerging role of the molecular marker p27 in the differential diagnosis of adrenocortical tumors

doi: 10.1530/EC-13-0025

Figure Lengend Snippet: Graphical representation of ROC curves with the respective area under the curve (AUC) to compare carcinoma (ACC) and total adenoma (ACAt) samples for the markers IGF2, p27, and Ki-67.

Article Snippet: The sections were incubated overnight at 4 °C with the appropriate diluted primary antibodies: rabbit anti-human MABs to p53 (453M-94; 1:100; Cell Marque, Rocklin, CA, USA), p21 (421M-14; 1:50; Cell Marque), p27 (427M-94; 1:500; Cell Marque), cyclin D1 (271R-14; 1:500; Cell Marque), Ki-67 (27R-14; 1:100; Cell Marque), and E-cadherin (246R-14; 1:200; Cell Marque) and rabbit anti-human polyclonal antibodies to β-catenin (424A-14; 1:500; Cell Marque).

Techniques:

Journal: Endocrine Connections

Article Title: The emerging role of the molecular marker p27 in the differential diagnosis of adrenocortical tumors

doi: 10.1530/EC-13-0025

Figure Lengend Snippet: Graphical representation of ROC curves with the respective area under the curve (AUC) to compare carcinoma (ACC) and nonfunctioning adenoma (ACAn) samples for the markers IGF2, p27, and Ki-67.

Article Snippet: The sections were incubated overnight at 4 °C with the appropriate diluted primary antibodies: rabbit anti-human MABs to p53 (453M-94; 1:100; Cell Marque, Rocklin, CA, USA), p21 (421M-14; 1:50; Cell Marque), p27 (427M-94; 1:500; Cell Marque), cyclin D1 (271R-14; 1:500; Cell Marque), Ki-67 (27R-14; 1:100; Cell Marque), and E-cadherin (246R-14; 1:200; Cell Marque) and rabbit anti-human polyclonal antibodies to β-catenin (424A-14; 1:500; Cell Marque).

Techniques: