Journal: Oncology reports

Article Title: Impact of Mucin1 knockdown on the phenotypic characteristics of the human hepatocellular carcinoma cell line SMMC-7721.

doi: 10.3892/or.2014.3136

Figure Lengend Snippet: Figure 3. Knockdown of MUC1 expression alters the β‑catenin signaling pathway by blocking β‑catenin translocation to the nucleus. (A) Cell lysates from NC and MR1-D4 clones were subjected to immunoprecipitation (IP) with anti-MUC1-CT antibody or normal IgG and then immunoblotted (IB) with anti‑β‑catenin antibody. Whole cell lysate (WCL) was not subjected to immunoprecipitation. (B) Cytoplasmic extracts from SMMC-7721, NC, MR1-C6 and MR1-D4 cells were analyzed by western blotting for the expression of β‑catenin. Cytoplasmic IκBα was used as a protein loading control. (C) Nuclear extracts from SMMC‑7721, NC, MR1-C6 and MR1-D4 cells were detected by western blotting to assess the levels of nuclear β‑catenin. Lamin B1 served as the nuclear loading control. (D) NC, MR1-C6 and MR1-D4 cells were transiently transfected with TOPflash and FOPflash plasmids. Relative luciferase activity was cal culated as the ratio of TOPflash/FOPflash luciferase activity, and each value was normalized to the luciferase activity of the internal control pRL-TK reporter plasmid. (E) mRNA levels of cyclin D1 and c-Myc in NC, MR1-C6 and MR1-D4 cells were detected by qRT-PCR and normalized to β‑actin. Bars represent the relative mRNA level when compared to the NC cells. (F) Cell lysates were analyzed by western blotting for the expression of cyclin D1 and c-Myc. β‑actin was used as a loading control. Data are expressed as the means ± SDs of 3 independent experiments. *P<0.05 compared with NC.

Article Snippet: The primary antibodies used were antibodies against MUC1 (GP1.4) (1:2,000; NeoMarkers), c-Myc (1:1,000), cyclin D1 (1:1,000), β-actin (1:2,000), IκBα (1:2,000) and Lamin B1 (1:2,000; all from Epitomics, Burlingame, CA, USA), β-catenin (1:1,000; BD Biosciences), E-cadherin (Proteintech), caspase-3 (Santa Cruz Biotechnology).

Techniques: Knockdown, Expressing, Blocking Assay, Translocation Assay, Clone Assay, Immunoprecipitation, Western Blot, Control, Transfection, Luciferase, Activity Assay, Plasmid Preparation, Quantitative RT-PCR

Journal: The EMBO journal

Article Title: Coordinated removal of repressive epigenetic modifications during induced reversal of cell identity.

doi: 10.15252/embj.2019101681

Figure Lengend Snippet: Figure 1. Kdm3b plays an important role in reprogramming to pluripotency.

Article Snippet: Antibodies used were H3K9me1 1:5,000 (Abcam Ab9045), H3K9me2 1:1,000 (Abcam Ab1220), Kdm3b 1:1,000 (Cell Signaling 5377), Kdm3a 1:1,000 (Proteintech, 12835-1-AP), 1:1,000 Jmjd1c (MBL, D356-3), 1:1,000 POU5F1 (Santa Cruz, sc-5279 or sc-8628), 1:1,000 alpha-Tubulin (Cell Signaling, 3873), and 1:1,000 NSD3 (Thermo Fisher, PA5-28972).

Techniques:

Journal: The EMBO journal

Article Title: Coordinated removal of repressive epigenetic modifications during induced reversal of cell identity.

doi: 10.15252/embj.2019101681

Figure Lengend Snippet: Figure 5. Kdm3b-KO cells retain 5hmC at pluripotency-associated locations where it should be resolved in reprogramming.

Article Snippet: Antibodies used were H3K9me1 1:5,000 (Abcam Ab9045), H3K9me2 1:1,000 (Abcam Ab1220), Kdm3b 1:1,000 (Cell Signaling 5377), Kdm3a 1:1,000 (Proteintech, 12835-1-AP), 1:1,000 Jmjd1c (MBL, D356-3), 1:1,000 POU5F1 (Santa Cruz, sc-5279 or sc-8628), 1:1,000 alpha-Tubulin (Cell Signaling, 3873), and 1:1,000 NSD3 (Thermo Fisher, PA5-28972).

Techniques:

Journal: bioRxiv

Article Title: Validation of GCN5L1/BLOC1S1/BLOS1 Antibodies Using Knockout Cells and Tissue

doi: 10.1101/2023.07.21.550091

Figure Lengend Snippet: Quantitative PCR (qPCR) was used to confirm that GCN5L1 gene expression was absent in KO MEF cells. N = 3.

Article Snippet: The peptide immunogens used to generate each of the GCN5L1 antibodies (other than the Proteintech antibody, which was not reported by the manufacturer) are shown in .

Techniques: Real-time Polymerase Chain Reaction, Gene Expression

Journal: bioRxiv

Article Title: Validation of GCN5L1/BLOC1S1/BLOS1 Antibodies Using Knockout Cells and Tissue

doi: 10.1101/2023.07.21.550091

Figure Lengend Snippet: The epitopes of each of the antibodies used in this study mapped to human GCN5L1 (1-153 aa). The peptide immunogen sequence for the Proteintech antibody was not reported.

Article Snippet: The peptide immunogens used to generate each of the GCN5L1 antibodies (other than the Proteintech antibody, which was not reported by the manufacturer) are shown in .

Techniques: Sequencing

Journal: bioRxiv

Article Title: Validation of GCN5L1/BLOC1S1/BLOS1 Antibodies Using Knockout Cells and Tissue

doi: 10.1101/2023.07.21.550091

Figure Lengend Snippet: GCN5L1 protein was essentially undetectable in KO MEFs using the Covance and Sigma antibodies (A,C). In contrast, an immunoreactive band at the correct molecular weight for GCN5L1 was detected in validated KO MEFs using both the Proteintech and Santa Cruz antibodies (B,D). N = 3.

Article Snippet: The peptide immunogens used to generate each of the GCN5L1 antibodies (other than the Proteintech antibody, which was not reported by the manufacturer) are shown in .

Techniques: Molecular Weight

Journal: bioRxiv

Article Title: Validation of GCN5L1/BLOC1S1/BLOS1 Antibodies Using Knockout Cells and Tissue

doi: 10.1101/2023.07.21.550091

Figure Lengend Snippet: Quantitative PCR (qPCR) was used to confirm that GCN5L1 gene expression was absent in KO liver tissue. N = 3

Article Snippet: The peptide immunogens used to generate each of the GCN5L1 antibodies (other than the Proteintech antibody, which was not reported by the manufacturer) are shown in .

Techniques: Real-time Polymerase Chain Reaction, Gene Expression

Journal: bioRxiv

Article Title: Validation of GCN5L1/BLOC1S1/BLOS1 Antibodies Using Knockout Cells and Tissue

doi: 10.1101/2023.07.21.550091

Figure Lengend Snippet: GCN5L1 protein was essentially undetectable in KO livers using the Covance and Sigma antibodies (A,C). In contrast, an immunoreactive band at the correct molecular weight for GCN5L1 was detected in validated KO livers using the Proteintech antibody (B). N = 3.

Article Snippet: The peptide immunogens used to generate each of the GCN5L1 antibodies (other than the Proteintech antibody, which was not reported by the manufacturer) are shown in .

Techniques: Molecular Weight

Journal: bioRxiv

Article Title: Validation of GCN5L1/BLOC1S1/BLOS1 Antibodies Using Knockout Cells and Tissue

doi: 10.1101/2023.07.21.550091

Figure Lengend Snippet: GCN5L1 protein was not detectable in either wildtype (WT) or knockout (KO) liver tissue using the Santa Cruz antibody. N = 3.

Article Snippet: The peptide immunogens used to generate each of the GCN5L1 antibodies (other than the Proteintech antibody, which was not reported by the manufacturer) are shown in .

Techniques: Knock-Out