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: Prostaglandins, leukotrienes, and essential fatty acids

Article Title: Fatty acid metabolism is involved in both retinal physiology and the pathology of retinal vascular diseases.

doi: 10.1016/j.plefa.2022.102473

Figure Lengend Snippet: Fig. 1. Representative immunofluorescent labeling of healthy human retina with several anti-FABPs. Paraffine-fixed healthy human retina was immunolabeled with anti-FABP3, FABP4, FABP5, FABP7 FABP8 or FABP12 as described in the methods section, and representative merged images of the immunolabeling with DAPI are shown. Scale bar; 25 µm. Experiments were repeated twice at a minimum to confirm that the staining is correct, and among these, representative images were used. FABP; fatty acid binding protein, NFL; nerve fiber layer, GCL; ganglion cell layer, IPL; inner plexiform layer, INL; inner nuclear layer, OPL; outer nuclear layer, ONL; outer nuclear layer, OS; outer segment, RPE; retinal pigment epithelium.

Article Snippet: After the deparaffinization, the sections were each blocked by treatment with 3% bovine serum albumin (BSA) for 1 hour, and successively incubated with the 1st antibody including anti-FABP3 (#10,676-AP, Proteintech, Rosemont, IL, USA), FABP4 (#13,979, Abcam, Eugene, OR, USA), FABP5 (#HP9030–100UG, Wayne, PA, USA), FABP7 (#51,010–1-AP, Proteintech, Rosemont, IL, USA), FABP8 (#12,717–1-AP, Proteintech, Rosemont, IL, USA) or FABP12 (#155,089, Abcam, Eugene, OR, USA) for 3 hrs, and the 2nd antibody; a goat anti-rabbit IgG (488 nm, 1:1000 dilutions) and DAPI (1:1000 dilutions) for 3 hrs.

Techniques: Labeling, Immunolabeling, Staining, Binding Assay

Journal: Prostaglandins, leukotrienes, and essential fatty acids

Article Title: Fatty acid metabolism is involved in both retinal physiology and the pathology of retinal vascular diseases.

doi: 10.1016/j.plefa.2022.102473

Figure Lengend Snippet: Fig. 2. Representative immunofluorescent labeling of wild type (WT), SDT and P23H rats and WT mouse retinas with several anti-FAPB4 antibodies. Paraffin-fixed Rat (WT, SDT or P23H) rat or WT mouse retina was immunolabeled with anti-FABP3, FABP4, FABP5, FABP7, FABP8 or FABP12 as described in the methods section. Representative immunolabeling images (left) and merged images with DAPI (right) of samples stained with FABP4 and FABP12 are shown, since no staining was observed by other FABP antibodies as shown in Table 1. Scale bar; 25 µm. Experiments were repeated twice at a minimum to confirm that the staining is correct, and among these, the representative images were used. FABP; fatty acid binding protein, NFL; nerve fiber layer, GCL; ganglion cell layer, IPL; inner plexiform layer, INL; inner nuclear layer, OPL; outer nuclear layer, ONL; outer nuclear layer, OS; outer segment, RPE; retinal pigment epithelium.

Article Snippet: After the deparaffinization, the sections were each blocked by treatment with 3% bovine serum albumin (BSA) for 1 hour, and successively incubated with the 1st antibody including anti-FABP3 (#10,676-AP, Proteintech, Rosemont, IL, USA), FABP4 (#13,979, Abcam, Eugene, OR, USA), FABP5 (#HP9030–100UG, Wayne, PA, USA), FABP7 (#51,010–1-AP, Proteintech, Rosemont, IL, USA), FABP8 (#12,717–1-AP, Proteintech, Rosemont, IL, USA) or FABP12 (#155,089, Abcam, Eugene, OR, USA) for 3 hrs, and the 2nd antibody; a goat anti-rabbit IgG (488 nm, 1:1000 dilutions) and DAPI (1:1000 dilutions) for 3 hrs.

Techniques: Labeling, Immunolabeling, Staining, Binding Assay