Figure 3.

Characteristics of SGOs and SGMCs derived from Cont‐ and OVX mice. a) Representative images of SGOs derived from Cont‐ and OVX mice at day 7. b) Total count and proportion of spheroid‐shaped organoids assessed at day 7. c) qPCR results showing the mRNA expression levels of Ck14, Ck19 and Tgfb2 in cont‐ and OVX‐SGOs at P0 and P2. d,e) qPCR results comparing the mRNA expression of Tgfb2 (d) and MCP genes (e) between SGMCs isolated from Cont‐ and OVX mice. f) Immunofluorescence detection of THBS1 and TGFβ2 expression in SGMCs. g) ELISA‐based measurement of TGFβ2 levels released into the SGMC medium. h,i) SGOs were cultured with CMs from Cont‐ and OVX‐SGMCs (Cont‐CM and OVX‐CM) (n = 2 for each) for 48 h, then the impact of SGMC‐CMs on SGO viability (h) and Smad2 activation (p‐SMAD2/SMAD2 ratio) (i) were determined by PI staining and WB, respectively. j) SGOs were cultured with SGMC‐CM derived from OVX mice with or without SB treatment (10 µm) for 48 h, then their viability was assessed by quantification of PI⁺ SGOs. At least three lines of SGOs, SGMCs, and their CMs were used for all experiments, except for (h) and (j), where CMs were collected from two lines of SGMCs. Scale bar = 500 µm (a, h, i) and 40 µm (f). Data are shown as the mean ± SEM and compared by One‐way ANOVA with Dunnett`s multiple comparisons tests (h) or unpaired t‐test (rest of the analysis). *P < 0.05, **P < 0.01, ***P < 0.001.