Journal: bioRxiv

Article Title: Effects of Alternative Splicing-Specific Knockdown of Tjp1 α+ by Rbm47 on Tight Junctions Assembly during Blastocyst Development

doi: 10.1101/2023.07.18.549609

Figure Lengend Snippet: (A) The position of both isoforms of Tjp1 (exon11/12; α+/α-), the α-domain (exon20/21) and two Tjp1 α-specific (exon 19/21; w/o α and alternatively spliced exon 27/28; α-) primers used for cDNA amplification of Tjp1 isoforms. Arrows mark the position and direction of primers used for cDNA synthesis. Transcription levels of Tjp1 α+/-, Tjp1 α variant (B), and Tjp1 α+/-, Tjp1 α-, Rbm47 (C). Transcription levels of Tjp1 α+/α-, Tjp1 α-, and Rbm47, shown as relative quantification (RQ). Error bars represent mean ± standard error.

Article Snippet: Primary antibodies against RBM47 (HPA006347, Sigma-Aldrich), TJP1 α+ (HP9044; Hycult Biotechnology, Uden, Netherlands) and TJP1 α-(HP9045; Hycult Biotechnology) were diluted and incubated with the embryos overnight at 4°C.

Techniques: Amplification, Variant Assay

Journal: bioRxiv

Article Title: Effects of Alternative Splicing-Specific Knockdown of Tjp1 α+ by Rbm47 on Tight Junctions Assembly during Blastocyst Development

doi: 10.1101/2023.07.18.549609

Figure Lengend Snippet: (A) Expression and localisation of Tjp1 α+ and Tjp1 α-in the Rbm47 KD blastocyst. (B) Gene expression patterns of Tjp1 variants from 8-cell to hatching blastocyst (C) Proximity ligation assay (PLA) showing the interaction between Tjp1(mainly Tjp1 α-) and Ocln in blastocysts. 8C(8-cell), 8c-cmp (compacted 8-cell), Mo (Morula), Lt-Mo (Late morula), E-Bl (early blastocyst), Ex-Bl (Expanding-Bl), H-Bl (Hatching blastocyst). Antibodies host: Rabbit (Rb), Guinea pig (GP), mouse (m).

Article Snippet: Primary antibodies against RBM47 (HPA006347, Sigma-Aldrich), TJP1 α+ (HP9044; Hycult Biotechnology, Uden, Netherlands) and TJP1 α-(HP9045; Hycult Biotechnology) were diluted and incubated with the embryos overnight at 4°C.

Techniques: Expressing, Proximity Ligation Assay

Journal: Cell Cycle

Article Title: Chibby1 knockdown promotes mesenchymal-to-epithelial transition-like changes

doi: 10.1080/15384101.2017.1281478

Figure Lengend Snippet: Cby1 KD results in a dramatic increase in ZO-1 protein levels at the plasma membrane. A. IF staining for the epithelial marker ZO-1 in control or Cby1-KD SW480 or HEK293 cells as indicated. Nuclei were visualized with DAPI. Scale bar, 20 μm. B. (Left) Cell lysates from control or Cby1-KD HEK293 cells were subjected to western blotting with ZO-1 and β-actin (loading control) antibodies. (Right) The band intensity of ZO-1 was quantified and normalized to that of β-actin. The results are expressed as mean ± SEM from triplicate samples in a single representative experiment. The control scrambled shRNA is set as 1. * P < 0.05.

Article Snippet: The primary antibodies used were as follows: mouse anti-β-catenin and mouse anti-E-cadherin (BD Transduction Laboratories); mouse anti-ZO-1 (BD Transduction Laboratories).

Techniques: Staining, Marker, Western Blot, shRNA