Journal: Cells

Article Title: A New Immortalized Human Lacrimal Gland Cell Line

doi: 10.3390/cells13070622

Figure Lengend Snippet: Primers RT-PCR.

Article Snippet: The cells were blocked in PBS and Tween-20 with 1% ( w / v ) nonfat dry milk for 1 h at RT followed by incubation with 1:50 PAX6 (AF8150; R&D Systems, Minneapolis, MN, USA), 1:500 FOXC1/2 (CPA3482; Cohesion, Suzhou, China), CLDN5 (MA532614, Invitrogen, Waltham, MA, USA), 1:50 KRT19 (M0888; Dako, Santa Clara, CA, USA) or OCLN (33-1500, Thermo Fisher Scientific, Waltham, MA, USA) primary antibody dilutions overnight at 4 °C in a humidified chamber.

Techniques: Sequencing

Journal: Cells

Article Title: A New Immortalized Human Lacrimal Gland Cell Line

doi: 10.3390/cells13070622

Figure Lengend Snippet: Antibodies for immunofluorescence staining.

Article Snippet: The cells were blocked in PBS and Tween-20 with 1% ( w / v ) nonfat dry milk for 1 h at RT followed by incubation with 1:50 PAX6 (AF8150; R&D Systems, Minneapolis, MN, USA), 1:500 FOXC1/2 (CPA3482; Cohesion, Suzhou, China), CLDN5 (MA532614, Invitrogen, Waltham, MA, USA), 1:50 KRT19 (M0888; Dako, Santa Clara, CA, USA) or OCLN (33-1500, Thermo Fisher Scientific, Waltham, MA, USA) primary antibody dilutions overnight at 4 °C in a humidified chamber.

Techniques: Immunofluorescence, Staining

Journal: Cells

Article Title: A New Immortalized Human Lacrimal Gland Cell Line

doi: 10.3390/cells13070622

Figure Lengend Snippet: Validation of lacrimal gland markers at the transcriptional and protein levels. ( A ) Gene expression of lacrimal gland epithelial marker genes in clones 1, 3 and 6 (RNA counts imply counts per million). A list of all gene abbreviations can be found in . ( B ) Agarose gels with the amplification products of RT-PCR against marker genes tested in clone 1 (C1), clone 3 (C3), clone 6 (C6), human fibroblasts (Fibro) and human corneal epithelial cells (CEC). A list of marker genes can be found in . ( C ) Semiquantitative comparison of gene expression after RT-PCR and gel electrophoresis. A list of marker genes can be found in . Data are means ± SEMs ** p < 0.01, *** p < 0.001, and **** p < 0.0001, two-way ANOVA, n = 6. ( D ) Immunofluorescence images of clones 1, 3 and 6 stained with DAPI (blue), Paired Box Gene 6 ( PAX6 , green, upper row) and Forkhead Box C1/C2 ( FOXC1/2 ), green, bottom row.

Article Snippet: The cells were blocked in PBS and Tween-20 with 1% ( w / v ) nonfat dry milk for 1 h at RT followed by incubation with 1:50 PAX6 (AF8150; R&D Systems, Minneapolis, MN, USA), 1:500 FOXC1/2 (CPA3482; Cohesion, Suzhou, China), CLDN5 (MA532614, Invitrogen, Waltham, MA, USA), 1:50 KRT19 (M0888; Dako, Santa Clara, CA, USA) or OCLN (33-1500, Thermo Fisher Scientific, Waltham, MA, USA) primary antibody dilutions overnight at 4 °C in a humidified chamber.

Techniques: Biomarker Discovery, Gene Expression, Marker, Clone Assay, Amplification, Reverse Transcription Polymerase Chain Reaction, Comparison, Nucleic Acid Electrophoresis, Immunofluorescence, Staining

Journal: International Journal of Molecular Sciences

Article Title: Gonadal, Not Maternal, Acquisition of Duplicated pax6 Orthologs in Megalobrama Amblycephala

doi: 10.3390/ijms20071710

Figure Lengend Snippet: Comparison of the two pax6 genes of Megalobrama amblycephala . ( A ) The genomic structures of Mapax6a and Mapax6b are shown. Conserved exons were connected by dotted lines. Both the translation-start sites (ATG) of Mapax6a and Mapax6b are located in exon 4. The mRNA structure is under the genomic structure. The paired domain is stained in blue, the green area is the homeodomain, and the yellow represents the proline–serine–threonine (PST)-rich transactivation domain. The comparison percentages of the open reading frame (ORF), 3′ untranslated region (UTR), and protein between Mapax6a and Mapax6b were analyzed by DNAMAN software. An alignment of the nucleic acid and amino acid sequence of exon 5a is under the mRNA structure (a/b). ( B ). Synteny conservation of pax6 and adjacent genes in different species. The duplicated pax6s are indicated in medaka and zebrafish. +: forward direction; -: reverse direction. ( C ) Summary isoforms of Mapax6a and Mapax6b . RNA-seq detected all of these isoforms, which were produced from alternative exons such as exon 2, exon3, and exon 5a.

Article Snippet: The membrane was blocked with 5% non-fat dry milk in Tris-Buffered Saline Tween-20 (TBST) for 1 h, then incubated with anti-Pax6 rabbit polyclonal antibody (Boster, Pleasanton, CA, USA, dilution 1:250) or actin antibody (Vazyme, Nanjing, China, dilution 1:1000) overnight at 4 °C.

Techniques: Comparison, Staining, Software, Sequencing, RNA Sequencing, Produced

Journal: International Journal of Molecular Sciences

Article Title: Gonadal, Not Maternal, Acquisition of Duplicated pax6 Orthologs in Megalobrama Amblycephala

doi: 10.3390/ijms20071710

Figure Lengend Snippet: Molecular phylogenetic analysis of pax6 homologs. The phylogenetic relationship of PAX family proteins was analyzed using the MEGA6 program by bootstrap analysis using neighbor-joining (1000 replicates). The numbers at the forks are the bootstrap proportions for each branch. A star indicates Mapax6a, and a triangle indicates Mapax6b.

Article Snippet: The membrane was blocked with 5% non-fat dry milk in Tris-Buffered Saline Tween-20 (TBST) for 1 h, then incubated with anti-Pax6 rabbit polyclonal antibody (Boster, Pleasanton, CA, USA, dilution 1:250) or actin antibody (Vazyme, Nanjing, China, dilution 1:1000) overnight at 4 °C.

Techniques: