Fig 2: CircTNRC18 inhibits Trophoblast Cell EMT by regulating EMT-related gene. (a) Western blot analysis was used to determine Grhl2, Vim and E-cadherin protein levels in HTR8/SVneo cells after transfecting with circTNRC18 overexpression vector or vector control. ß-Actin was used as an internal control. Densitometric analysed the three independent experiments. **P < 0.01 vs. vector control. (b) HTR8/SVneo cells were transfected with si-circTNRC18-1#, si-circTNRC18-2#, or negative control (si-NC). Western blot analysis was used to examine Grhl2, Vim, E-cadherin protein levels.**P < 0.01, ***P < 0.001 vs. si-NC. (c and d) HTR8/SVneo cells were prepared as in (a) and (b). Expressions of E-cadherin, Vim and ß-catenin were examined by Immunofluorescence staining. Scale bars = 20 µm.

Fig 3: In vitro and in vivo images of hVM1 clone 32 cell markers. (A) After 7 days of differentiation in vitro, hVM1 clone 32 cells express a range of proteins including TH, TUBB3, Ki-67, VIM, MAP2, SYN1, GFAP, and GABA (all in white). Nuclei were stained with DAPI (blue). Scale bar = 20 µm. (B) Surviving transplanted hNSCs in the Str 4 months post-transplant as marked by STEM121 in red which stains human-specific cytoplasm (left), and astrocytes as marked by GFAP in grey near the transplanted cells (right). Nuclei were stained with DAPI (blue). Scale bars = 100 µm.

Fig 4: IBA1+-cells express CD163, a marker for border-associated macrophagesImmunostaining of cryostat sections of SC102A 3D-cysts from unguided protocol for IBA1 (ionized calcium-binding adapter molecule 1, magenta) and the nuclei-dye Hoechst (blue). *, lumen within the cystic compartment.(A) Immunostaining of CD163 (cluster of differentiation 163/protein tyrosine phosphatase receptor type C, green) in SC102A at week 8 with zoom-in (Orange frame). Scale bar: 100 µm.(B) Bar chart of % of CD163+/IBA1+-cells and % of CD163-/IBA1+-cells per time point with SE. Each dot represents one section of an individual 3D-cyst. two-way ANOVA p value = < 2.2e-16 with selected post hoc test. ***p < 0.001.(C and D) CD163 expression in 2.5D-culture for CR05 at week 5 (C) and week 6 (D). Open arrow, CD163-/IBA1+. Arrow, CD163+/IBA1+. Red frame, zoom in. Scale bar: 100 µm.(E) Immunostaining for CD31 (platelet and endothelial cell adhesion molecule 1, green) and VIM (vimentin, yellow) for SC102A at week 8. Scale bar: 100 µm. Zoom-in, 3D-surface rendering (i, ii). Scale bar: 20 µm.

Fig 5: NR morphology immunochemical evaluation. Immunoreactivity of PNA lectin (A–D), rho protein (E–H), PKC a isoform (PKCa) (I–L), NeuN (M–P), GFAP (Q–T) and Vim (U–X). The experimental conditions analyzed were fresh neuroretinas (Fresh), mono-cultured NR explants for 72 hours (NR), NR explants co-cultured with human bmMSC for 72 hours (NR + MSC), and NR explants cultured with medium from NR + MSC co-culture for 72 hours (NR + S). Scale bar: 25 µm.