WFS1 deficiency impacts GnRH development through ER stress response failure

Supporting Data Gentile et al (excel file): this dataset support the involvment of wfs1 in gnrh neurons develpment. This dataset provides alla raw numberical data corrisponding to the graphs presented in the main text figures and supplmentary materials of this study. Depending on the experiments, statistics were calculated using Student-T test and One way Anova using GraphPad Prism 9 software. Each subfolder is named according to the corresponding figure, as follows: FIG.1I: quantification of optical density of western blots bands using ImageJ software. The intensity values of WFS1 plasmids are normalized to that of the beta actin. Results are expressed as mean ± SEM of three independent experiments. FIG.1J: in vitro luciferase assay expressed in RLU (Relative Light Units). Results are expressed as mean ± SEM of three independent experiments. FIG.1K-Q: quantification of the expression of BIP, ATF6, IRE1, PERK, ATF4, XBP1 and CHOP mRNAs by RT-qPCRcells transfected with wild type or mutated WFS1 plasmids. ddCT method was used to analyze the results, expressed as mean ± SEM from three independent experiments. Results were normalized with GAPDH as internal control and considering transcript levels of WT as 1. FIG.2D-E: quantification of the pixel area expressing the green fluorescence protein in different brain regions: Panel D (OBs and AC) and Panel E (POA, Re and Hy). Quantifications were obtained using ImageJ software. Results are expressed as mean ± SEM of t at least 10 embryos derived from three independent experiments. FIG.5A-G: quantification of the expression of bip, atf6, ire1, perk, atf4, xbp1 and chop mRNAs by RT-qPCR in pools of embryos at 48hpf injected with stdMO or wfs1bMOs in basal condition or treated with 5uM VPA. ddCT method was used to analyze the results, expressed as mean ± SEM from three independent experiments. Results were normalized with eef1a1 as internal control and considering transcript levels of stdMO as 1. FIG.6E: quantification of the axon length (um) in olfactory and hindbrain regions of stdMO, wfs1bMOs and isl1aMO at 48hpf using ImageJ software with SNT plugin. Results are expressed as mean ± SEM of t at least 12 embryos derived from three independent experiments. FIG.S2C: frequency of injected embryos presented normal, moderate and severe morphology at 48hpf. Results are expressed as mean ± SEM of t at least 250 embryos derived from five independent experiments.