Journal: iScience

Article Title: Iloprost requires the Frizzled-9 receptor to prevent lung cancer

doi: 10.1016/j.isci.2022.104442

Figure Lengend Snippet:

Article Snippet: Vectastain Elite ABC anti-mouse/Rabbit RTU HRP immunodetection kit , Novus Biologicals , PK-7200.

Techniques: Recombinant, Western Blot, Plasmid Preparation, Electrophoresis, Reporter Assay, BIA-KA, Reverse Transcription, Immunodetection, Knock-Out, Transgenic Assay, Software

Journal: Biology Open

Article Title: Defects in GPI biosynthesis perturb Cripto signaling during forebrain development in two new mouse models of holoprosencephaly

doi: 10.1242/bio.20121982

Figure Lengend Snippet: Wildtype ( A–C ) and gnz ( D–F ) mutant E18.5 embryos with lateral (B,E) and top (C,F) views of skull stained for bone (red) and cartilage (blue). Exoccipital (eo), supraoccipital (so), basioccipital (bo), tectum synoticum (tso) and interparietal (ip) elements are indicated. Critical 5.14 Mb region of chromosome 1 that the gnz mutation mapped to by meiotic recombination ( G ). Recombinants are shown in parentheses divided by number of recombination opportunities. SNP and SSLP markers and known or predicted genes (horizontal bars) are indicated. There is a T to A transversion in the splice donor of Pign's intron 23 ( H ) that causes skipping of exons 23 and 24 ( J ) as shown by PCR amplification of wildtype (WT), heterozygote (Het) and gnz mutant cDNA ( I ). Mis-splicing of exon 22 to 25 results in a frame shift and premature stop codon within 13 amino acids from the mutation ( K ) leading to truncation and partial loss of the catalytic domain and KKXX ER-retention motif ( L ). Pign mRNA detected by whole mount in situ hybridization in E14.5 wildtype ( M ) and Pign gnz/gnz mutant ( N ) embryos, showing ubiquitous and stable expression in gnz mutants. GFP fluorescence was detected in E13.5 Pign +/+ ; Tg GPI-GFP ( O -left) but was greatly reduced in Pign gnz/gnz ; Tg GPI-GFP (O -right) embryos and MEFs ( P–S ; P and R are Pign +/+ ; Tg GPI-GFP MEFs and Q and S are Pign gnz/gnz ; Tg GPI-GFP MEFs). Panels R and S were immunostained for the Golgi marker βCOP (red). See also supplementary material Fig. S1 for mutant phenotypes. Ratios in lower right of panels N,O and subsequent figures indicate number of similar phenotypes observed out of number of mutant embryos analyzed. Scale bars (A–F) = 2 mm.

Article Snippet: MEFs were grown in 10% FBS in DMEM, fixed in 4% paraformaldehyde then analyzed by immunodetection for βCOP (GeneTex, Inc).

Techniques: Mutagenesis, Staining, Amplification, In Situ Hybridization, Expressing, Fluorescence, Marker