Journal: Nature Communications
Article Title: Identifying fibroblast-derived sFRP2 as a therapeutic target and engineering siRNA therapy for uterine scarring
doi: 10.1038/s41467-025-62248-1
Figure Lengend Snippet: a Venn diagram analysis of differentially expressed WNT family genes comparing human (blue) and mouse (red) transcriptomes. b qPCR analysis of the expression of WNT16, WNT5A, and WNT9A in human myometrial and mouse uterine scar tissues. c WB analysis of WNT5A levels in human myometrial tissues. d Immunofluorescence imaging and plot profile analysis delineate the co-localization of sFRP2 with WNT5A in the human myometrium with US ( n = 6 samples). e , f WB ( e ) and immunofluorescence ( f ) analyses of WNT5A in hUFIBs ( n = 6 samples). g WB analysis of WNT5A in hUFIBs following sFRP2 overexpression (sFRP2OE), PBS-treated hUFIBs (WT), and negative control lentivector-treated hUFIBs (Control). h WB bands illustrate WNT5A levels in sFRP2-knockdown hUFIBs with TGF-β. WT cells received PBS and TGF-β, while control cells were treated with scramble shRNA and TGF-β. i WNT5A concentrations in conditioned media (CM) derived from WT, Control, and sFRP2OE groups, as well as corresponding heparinase-supplemented counterparts. j Immunoprecipitation (IP) assay indicates the interaction between FLAG-sFRP2 and HA-WNT5A in normal hUFIBs. k – m WB ( k , l ) and immunofluorescence ( m , n = 6 samples) analyses of fibrinogen, Col1A1, and Col3A1 in hUFIBs after WNT5A stimulation, using PBS as the control treatment. n , o Immunofluorescence ( n , n = 6 samples) and flow cytometric ( o ) analyses illustrate Ca 2+ influx (stained with Fluro-4 AM) in hUFIBs. p Flow cytometric quantification of Ca 2+ influx in hUFIBs subjected to different treatments. q Immunofluorescence analysis of the relationship of sFRP2 with Ca 2+ and fibrinogen in sFRP2OE hUFIBs ( n = 6 samples). r Dynamic tracing of Ca 2+ influx in hUFIBs of sFRP2KD, WT, and Control groups with TGF-β ( n = 6 samples). Box plots in b , i , l , o , p illustrate data distributions ( n = 6 samples), with the central lines representing the mean, box boundaries indicating standard deviation, and whiskers showing the minimum and maximum values. The p values were calculated by two-tailed unpaired t -tests for two groups and one-way ANOVA with post hoc LSD tests for multiple groups. Source data are provided as a Source Data file.
Article Snippet: For immunoprecipitation assay, 500 μg of total protein from cell lysis in each group (Control, HA-WNT5A-transfected cells, FLAG-sFRP2-transfected cells, or HA-WNT5A/FLAG-sFRP2-cotransfected cells) was incubated with 20 μL of anti-HA magnetic beads (MedChemExpress, HY-K0237) or anti-FLAG magnetic beads (MedChemExpress, HY-K0207) at 4 °C with gentle rotation overnight.
Techniques: Expressing, Immunofluorescence, Imaging, Over Expression, Negative Control, Control, Knockdown, shRNA, Derivative Assay, Immunoprecipitation, Staining, Standard Deviation, Two Tailed Test
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