Journal: BMC Biology

Article Title: Grb7 , Grb10 and Grb14, encoding the growth factor receptor-bound 7 family of signalling adaptor proteins have overlapping functions in the regulation of fetal growth and post-natal glucose metabolism

doi: 10.1186/s12915-024-02018-5

Figure Lengend Snippet: Expression of Grb7 family members in PN21 mouse pancreas visualised by immunohistochemistry on paraffin sections. Antibodies specific for ( A ) Grb7; B Grb10; and C Grb14; were used to detect distinct proteins of the Grb7 family. Brown staining is indicative of protein expression. Arrows indicate endocrine area (pancreatic islets); magnification 100x

Article Snippet: For immunohistochemistry, protocols were essentially as previously described [ ], using primary antibodies specific for either Grb7 (Santa Cruz Biotechnology, CA, USA; GRB7 (N-20) N-terminal Grb7, rabbit polyclonal sc-607; RRID:AB_2113275), at 1:200 dilution, Grb10 (Santa Cruz Biotechnology, CA, USA; GRB10 (A-18) C-terminal α-Grb10, rabbit polyclonal sc-1027) at 1:200 dilution or Grb14 (Santa Cruz Biotechnology, CA, USA; GRB14 (N-19) N-terminal Grb14, goat polyclonal sc-6103; RRID:AB_2112989) at 1:100 dilution.

Techniques: Expressing, Immunohistochemistry, Staining

Journal: BMC Biology

Article Title: Grb7 , Grb10 and Grb14, encoding the growth factor receptor-bound 7 family of signalling adaptor proteins have overlapping functions in the regulation of fetal growth and post-natal glucose metabolism

doi: 10.1186/s12915-024-02018-5

Figure Lengend Snippet: Weights and blood glucose levels in PN1 progeny from Grb10 KO x Grb14 KO crosses. Weights of whole body and selected dissected organs, with blood glucose levels were collected at PN1 from progeny of crosses between Grb10 KO and Grb14 KO mice. Data were pooled into four groups for analysis as described in the Methods; wild type (WT), Grb10 KO (10KO), Grb14 KO (14KO) and Grb10 : Grb14 double knockout (DKO). For each of the four offspring genotype groups, data are shown for ( A ) Body weight; and B Blood glucose concentration ([Glu]). In addition, actual weights of ( C ) Brain; D Liver; E Lungs; F Heart; and G Kidneys; are shown above the relative weights of the same organs, expressed as a percentage of body mass ( H – L ). Values represent means and SEM, tested by one-way ANOVA using Kruskal–Wallis and Dunn’s post hoc statistical tests. Sample sizes were, for total body and all organs, WT N = 25, Grb10 KO N = 13, Grb14 KO N = 7, Grb10 : Grb14 DKO N = 7; glucose levels, WT N = 18, Grb10 KO N = 9, Grb14 KO N = 6, Grb10 : Grb14 DKO N = 4. Asterisks indicate p- values, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: For immunohistochemistry, protocols were essentially as previously described [ ], using primary antibodies specific for either Grb7 (Santa Cruz Biotechnology, CA, USA; GRB7 (N-20) N-terminal Grb7, rabbit polyclonal sc-607; RRID:AB_2113275), at 1:200 dilution, Grb10 (Santa Cruz Biotechnology, CA, USA; GRB10 (A-18) C-terminal α-Grb10, rabbit polyclonal sc-1027) at 1:200 dilution or Grb14 (Santa Cruz Biotechnology, CA, USA; GRB14 (N-19) N-terminal Grb14, goat polyclonal sc-6103; RRID:AB_2112989) at 1:100 dilution.

Techniques: Double Knockout, Concentration Assay

Journal: BMC Biology

Article Title: Grb7 , Grb10 and Grb14, encoding the growth factor receptor-bound 7 family of signalling adaptor proteins have overlapping functions in the regulation of fetal growth and post-natal glucose metabolism

doi: 10.1186/s12915-024-02018-5

Figure Lengend Snippet: Genetic crosses used in the study, showing parent and offspring genotypes with their expected ratios. Crosses between (A) Grb10 +/p : Grb14 +/- double heterozygous females and Grb10 +/p : Grb14 +/- double heterozygous males, producing offspring of twelve genotypes, and (B) Grb7 +/- : Grb10 +/p double heterozygous females and Grb7 +/- heterozygous males, producing offspring of six genotypes, each in the indicated, expected Mendelian ratios. For ANOVA statistical analysis these six or twelve genotypes were used to form four groups, as indicated, since no difference was anticipated between animals differing only in their Grb14 +/- and Grb14 +/+ or Grb7 +/- and Grb7 +/+ allelic status. Similarly, due to the imprinted expression of Grb10 , offspring inheriting a mutant copy of the paternal Grb10 allele ( Grb10 +/p ), having no growth phenotype, were not expected to differ from Grb10 wild type ( Grb10 +/+ ) offspring, while double knockout (DKO) offspring inheriting mutations of both parental alleles ( Grb10 m/p ) were expected to be indistinguishable from those inheriting a mutant copy of the normally active maternal Grb10 allele ( Grb10 m/+ )

Article Snippet: For immunohistochemistry, protocols were essentially as previously described [ ], using primary antibodies specific for either Grb7 (Santa Cruz Biotechnology, CA, USA; GRB7 (N-20) N-terminal Grb7, rabbit polyclonal sc-607; RRID:AB_2113275), at 1:200 dilution, Grb10 (Santa Cruz Biotechnology, CA, USA; GRB10 (A-18) C-terminal α-Grb10, rabbit polyclonal sc-1027) at 1:200 dilution or Grb14 (Santa Cruz Biotechnology, CA, USA; GRB14 (N-19) N-terminal Grb14, goat polyclonal sc-6103; RRID:AB_2112989) at 1:100 dilution.

Techniques: Expressing, Mutagenesis, Double Knockout

Journal: BMC Biology

Article Title: Grb7 , Grb10 and Grb14, encoding the growth factor receptor-bound 7 family of signalling adaptor proteins have overlapping functions in the regulation of fetal growth and post-natal glucose metabolism

doi: 10.1186/s12915-024-02018-5

Figure Lengend Snippet: Summary of PN1 body and organ weight data for progeny of crosses between (A) the Grb10 KO and Grb14 KO strains, and (B) the Grb7 KO and Grb10 KO strains. Mean weights are shown for each genotype together with changes relative to wild type (%WT) for each mutant genotype

Article Snippet: For immunohistochemistry, protocols were essentially as previously described [ ], using primary antibodies specific for either Grb7 (Santa Cruz Biotechnology, CA, USA; GRB7 (N-20) N-terminal Grb7, rabbit polyclonal sc-607; RRID:AB_2113275), at 1:200 dilution, Grb10 (Santa Cruz Biotechnology, CA, USA; GRB10 (A-18) C-terminal α-Grb10, rabbit polyclonal sc-1027) at 1:200 dilution or Grb14 (Santa Cruz Biotechnology, CA, USA; GRB14 (N-19) N-terminal Grb14, goat polyclonal sc-6103; RRID:AB_2112989) at 1:100 dilution.

Techniques: Mutagenesis

Journal: BMC Biology

Article Title: Grb7 , Grb10 and Grb14, encoding the growth factor receptor-bound 7 family of signalling adaptor proteins have overlapping functions in the regulation of fetal growth and post-natal glucose metabolism

doi: 10.1186/s12915-024-02018-5

Figure Lengend Snippet: Weights and blood glucose levels in PN1 progeny from Grb7 KO x Grb10 KO crosses. Weights of whole body and selected dissected organs, with blood glucose levels were collected at PN1 from progeny of crosses between Grb7 KO and Grb10 KO mice. Data were pooled into four groups for analysis as described in the Methods; wild type (WT), Grb7 KO (7KO), Grb10 KO (10KO) and Grb7 : Grb10 double knockout (DKO). For each of the four offspring genotype groups, data are shown for, A Body weight; and B Blood glucose concentration ([glu]). In addition, actual weights of ( C ) Brain; D Liver; E Lungs; F Heart and G Kidneys are shown above the relative weights of the same organs, expressed as a percentage of body mass ( H – L ). Values represent means and SEM, tested by one-way ANOVA using Kruskal–Wallis and Dunn’s post hoc statistical tests. Sample sizes were, for body and brain, WT N = 42, Grb7 KO N = 14, Grb10 KO N = 49, Grb7 : Grb10 DKO N = 15; kidneys and heart, WT N = 42, Grb7 KO N = 14, Grb10 KO N = 48, Grb7 : Grb10 DKO N = 15, liver, WT N = 42, Grb7 KO N = 14, Grb10 KO N = 49, Grb7 : Grb10 DKO N = 14; lungs, WT N = 41, Grb7 KO N = 14, Grb10 KO N = 49, Grb7 : Grb10 DKO N = 15; glucose levels, WT N = 24, Grb7 KO N = 7, Grb10 KO N = 27, Grb7 : Grb10 DKO N = 10. Asterisks indicate p- values, * p < 0.05, ** p < 0.01, **** p < 0.0001

Article Snippet: For immunohistochemistry, protocols were essentially as previously described [ ], using primary antibodies specific for either Grb7 (Santa Cruz Biotechnology, CA, USA; GRB7 (N-20) N-terminal Grb7, rabbit polyclonal sc-607; RRID:AB_2113275), at 1:200 dilution, Grb10 (Santa Cruz Biotechnology, CA, USA; GRB10 (A-18) C-terminal α-Grb10, rabbit polyclonal sc-1027) at 1:200 dilution or Grb14 (Santa Cruz Biotechnology, CA, USA; GRB14 (N-19) N-terminal Grb14, goat polyclonal sc-6103; RRID:AB_2112989) at 1:100 dilution.

Techniques: Double Knockout, Concentration Assay

Journal: BMC Biology

Article Title: Grb7 , Grb10 and Grb14, encoding the growth factor receptor-bound 7 family of signalling adaptor proteins have overlapping functions in the regulation of fetal growth and post-natal glucose metabolism

doi: 10.1186/s12915-024-02018-5

Figure Lengend Snippet: Weight analysis of e17.5 conceptuses from crosses between Grb7 KO and Grb10 KO mice. Data were pooled into four groups for analysis as described in the Methods; wild type (WT), Grb7 KO (7KO), Grb10 KO (10KO) and Grb7 : Grb10 double knockouts (DKO). Weights are shown for the four offspring genotype groups for ( A ) Embryo; and B Placenta. C These values have been used to calculate the embryo to placenta weight ratio as a measure of placental efficiency. Values represent means and SEM, tested by one-way ANOVA using Kruskal–Wallis and Dunn’s post hoc statistical tests. Sample sizes were, for embryo WT N = 24, Grb7 KO N = 5, Grb10 KO N = 27, Grb7 : Grb10 DKO N = 6, and for placenta and embryo:placenta ratio WT N = 23, Grb7 KO N = 5, Grb10 KO N = 27, Grb7:Grb10 DKO N = 6. Asterisks indicate p- values, * p < 0.05, ** p < 0.01, **** p < 0.0001

Article Snippet: For immunohistochemistry, protocols were essentially as previously described [ ], using primary antibodies specific for either Grb7 (Santa Cruz Biotechnology, CA, USA; GRB7 (N-20) N-terminal Grb7, rabbit polyclonal sc-607; RRID:AB_2113275), at 1:200 dilution, Grb10 (Santa Cruz Biotechnology, CA, USA; GRB10 (A-18) C-terminal α-Grb10, rabbit polyclonal sc-1027) at 1:200 dilution or Grb14 (Santa Cruz Biotechnology, CA, USA; GRB14 (N-19) N-terminal Grb14, goat polyclonal sc-6103; RRID:AB_2112989) at 1:100 dilution.

Techniques:

Journal: BMC Biology

Article Title: Grb7 , Grb10 and Grb14, encoding the growth factor receptor-bound 7 family of signalling adaptor proteins have overlapping functions in the regulation of fetal growth and post-natal glucose metabolism

doi: 10.1186/s12915-024-02018-5

Figure Lengend Snippet: Grb7, Grb10 and Grb14 distribution in the developing mouse embryo

Article Snippet: For immunohistochemistry, protocols were essentially as previously described [ ], using primary antibodies specific for either Grb7 (Santa Cruz Biotechnology, CA, USA; GRB7 (N-20) N-terminal Grb7, rabbit polyclonal sc-607; RRID:AB_2113275), at 1:200 dilution, Grb10 (Santa Cruz Biotechnology, CA, USA; GRB10 (A-18) C-terminal α-Grb10, rabbit polyclonal sc-1027) at 1:200 dilution or Grb14 (Santa Cruz Biotechnology, CA, USA; GRB14 (N-19) N-terminal Grb14, goat polyclonal sc-6103; RRID:AB_2112989) at 1:100 dilution.

Techniques: