goat anti igf1  (R&D Systems)

Journal: Nature Communications

Article Title: A cell atlas of the human fallopian tube throughout the menstrual cycle and menopause

doi: 10.1038/s41467-024-55440-2

Figure Lengend Snippet: A Differences in cellular composition between the proliferative- ( n = 4) and secretory phases ( n = 3) of the menstrual cycle based on scRNA-seq data (two-sided t -test). Data are represented using a boxplot showing the median and first and third quantile. N describes the number of patients/ samples for each respective menstrual cycle phase. SE1 p-value < 0.0018, SE2 p -value < 0.0035. B Representative OVGP1 immunohistochemistry in the fallopian tube of patients in the proliferative phase ( n = 2) and in the secretory phase ( n = 2). C Scatter plot comparing gene expression levels of pre-menopausal SE1 (secretory phase) and SE2 cells (proliferative phase) using pseudo-bulk RNA analysis. D Volcano plot derived from pseudo bulk analysis. The volcano plot shows the differential gene expression analysis of genes expressed in SE cells (SE 1/2/3-pre) based on the proliferative and secretory phases. E Number of differentially expressed genes between the proliferative- and secretory phases of the menstrual cycle by cell clusters based on scRNA-seq data. F Dot plot showing normalized scRNA-seq derived gene expression levels of selected genes that differ in the proliferative and secretory phase in ST subtypes. G Representative immunofluorescence staining of IGF1 and IGF2 in primary human stromal and epithelial fallopian cell co-culture of one patient ( n = 3). The co-culture was stimulated for 8 h with estrogen (E4) or progesterone (P4). IGF1 protein in green, IGF2 protein in red and nuclei/ DAPI in blue. White dotted lines mark epithelial cells while white stars mark stromal cells. H Ligand-receptor interactions, detected by CellPhoneDB, between SE and ST subtypes (left) and between SE cells (right) separated by the menstrual phase using scRNA-seq data. I RNA velocity analysis of SE1- and SE2-pre cells. Arrows indicate the location of the estimated future cell state. Long vectors mark rapid transition events (i.e., large changes in gene expression), while short arrows indicate homeostasis. J Median latent time for pre-menopausal SE cells during the proliferative- and secretory phase of the menstrual cycle, highlighting temporal positions for SE1- and SE2-pre.

Article Snippet: Slides were stained overnight at 4 °C in goat serum (Thermo Fisher Scientific, Cat#16210064) using the following primary antibodies: IGF1 (1:200, OriGene Technologies, TA805748S) and IGF2 (1:200, ThermoFisher scientific, MA532485), followed by fluorescently labeled secondary antibodies (1:200, Alexa Fluor 488 and 568, Invitrogen) and Hoechst 33258 (1:200, Molecular probes, H-3569) for 1 h. We imaged slides on a Nikon Eclipse Ti2 microscope and processed images with NIS-Elements (Nikon).

Techniques: Immunohistochemistry, Gene Expression, Derivative Assay, Immunofluorescence, Staining, Co-Culture Assay