anti snail2  (Developmental Studies Hybridoma Bank)

Journal: Developmental cell

Article Title: Differential tissue deformability underlies fluid pressure-driven shape divergence of the avian embryonic brain and spinal cord.

doi: 10.1016/j.devcel.2025.04.010

Figure Lengend Snippet: Figure 3. Differential neural crest behavior underlies the acquisition of a thinned-out and expanded dorsal hindbrain roof (A) Confocal images of actin and phosphorylated myosin light chain kinase II in the hindbrain and spinal cord of HH12 stage embryos. (B and C) Quantification of apical actin signal intensity along the lumen circumference in the hindbrain and spinal cord of HH11-HH12 stage embryos (n = 3) (p = 0.012 and 0.53, paired t tests). (D) Confocal images showing Snail2+ cell migration in the hindbrain and spinal cord cross sections of embryos progressing toward brain-expansion stages (HH9 to HH12). (E) Confocal images showing laminin and actin organization in cross-section views of the hindbrain and spinal cord of HH12 stage embryos. (F) Confocal images showing laminin organization at the dorsal surface in a 3D rendering of the hindbrain and spinal cord of HH11 stage embryos. (G) Bright-field images of embryos treated with dimethyl sulfoxide (DMSO) or MMP inhibitor at HH11 and incubated for 20 h. Arrows indicate hindbrain at level of otic vesicle. (H) Dorsal views of DMSO- and MMP inhibitor-treated embryos. Yellow highlight fills the brain dorsal surface. (I) Confocal images showing the dorsal hindbrain of control and MMP inhibitor-treated embryos, ∼20 h post treatment. Bottom panels show laminin surface with arrows indicating breaks in laminin continuity. (J) Hindbrain roof length in control (n = 6) and inhibitor-treated (n = 8) embryos (p = 0.0000048, t test). White scale bars are 50 μm unless otherwise stated. See also Figure S3.

Article Snippet: The following antibodies and dyes where used: Laminin (DSHB, 3H11) 1:100, Snail2 (Cell Signaling Technology, 9585S) 1:200, pMLC2 (Cell Signaling Technology, 3671S) 1:100, Hoechst 1:1000, Phalloidin 647 (Thermo Scientific, A22287) 1:500, Donkey Anti-Mouse 488n IgG (Abcam, ab150105), Donkey Anti-Rabbit 594 (Abcam, ab150076).

Techniques: Migration, Incubation, Control

Journal: Developmental cell

Article Title: Differential tissue deformability underlies fluid pressure-driven shape divergence of the avian embryonic brain and spinal cord.

doi: 10.1016/j.devcel.2025.04.010

Figure Lengend Snippet: Figure 4. Hindbrain premigratory neural crest cells may be sufficient to generate neural tube expansion under lumen pres- sure (A) Bright-field images of pre-brain-expansion stage embryos with spinal cord-to-spinal cord graft and a hindbrain-to-spinal cord graft. (B and C) Confocal images showing dorsal surface of graft integration ∼20 h post-grafting and cross- sectional views at the levels corresponding to dashed lines in (B). White bracket highlights the thinned-out roof in the graft region. (D) Spinal cord tissue thickness in the region with spinal cord (n = 2) or hindbrain (n = 9) grafted cells. (E) Confocal images showing Snail2+ cells in graft regions. Arrows indicate Snail2+ cells within the GFP+ graft. (F) Model of brain expansion relative to the spinal cord. A greater extent of premigratory neural crest cell mesenchymal behavior and corresponding ECM remodeling underlies a more deformable dorsal roof in the early hindbrain compared with the spinal cord. This allows the hindbrain roof to deform more under internal lumen pressure, driving hindbrain expansion relative to the spinal cord during early embryo development. Black scale bars are 500 μm. White scale bars are 25 μm unless otherwise stated. See also Figure S4.

Article Snippet: The following antibodies and dyes where used: Laminin (DSHB, 3H11) 1:100, Snail2 (Cell Signaling Technology, 9585S) 1:200, pMLC2 (Cell Signaling Technology, 3671S) 1:100, Hoechst 1:1000, Phalloidin 647 (Thermo Scientific, A22287) 1:500, Donkey Anti-Mouse 488n IgG (Abcam, ab150105), Donkey Anti-Rabbit 594 (Abcam, ab150076).

Techniques:

Journal: International Journal of Medical Sciences

Article Title: Platelet-rich fibrin promotes mesothelial cell proliferation and peritoneal repair by up-regulating calretinin to prevent postoperative intestinal adhesion

doi: 10.7150/ijms.105523

Figure Lengend Snippet: Figure 6. PRF had a limited regulatory effect on the MMT of mesothelial cells. (A) The expression levels of E-cadherin, vimentin, and Snail2 in mesothelial cells treated with PRF or not. (B) Quantitative analysis results showed no significant difference in the protein levels of E-cadherin, vimentin, and Snail2 between the PRF group and control group (P > 0.05).

Article Snippet: The primary antibodies used include E-cadherin (Proteintech, 20874-1-AP, 1:1000 diluted, USA), vimentin (Bioss, bs-0756R, 1:1000 diluted, China), Snail2 (Proteintech, 12129-1-AP, 1:1000 diluted, USA), and the secondary antibodies used include Goal anti-rabbit antibody (680 anti-rabbit IgG, Yeasen, 33118ES60, 1:25000 diluted, China) and Goal anti-mouse antibody (790 anti-mouse IgG, Yeasen, 33219ES60, 1:25000 diluted, China).

Techniques: Expressing, Control

Journal: Differentiation; research in biological diversity

Article Title: Delamination of chick cephalic neural crest cells requires an MMP14-dependent downregulation of Cadherin-6B.

doi: 10.1016/j.diff.2025.100836

Figure Lengend Snippet: Fig. 2. MMP14 is required for the delamination of cephalic neural crest cells, independently of its catalytic activity. A, protocol of the Cornish pasty culture assay. B- C, transversal section at rhombomere 4 (r4) and rhombomere 6 (r6) level in embryos treated with DMSO (B) or NSC405020 (C, 0.5 mM). D, normalized area of the snail2-positive domain in the dorsal neural tube at antero-posterior levels and experimental conditions depicted in B-C (DMSO, nr4 = 4; nr6 = 4; NSC405020, nr4 = 7; nr6 = 7); T-test r4 DMSO vs NSC, p = 0.0061 (**), T-test r6 DMSO vs NSC, p = 0.23 (ns). E, Medio-lateral migration of snail2-positive neural crest cells at antero- posterior levels and experimental conditions depicted in B-C (DMSO, nr4 = 9; nr6 = 11; NSC, nr4 = 8; nr6 = 11); T-test r4 DMSO vs NSC, p = 0.0058 (**), T-test r6 DMSO vs NSC, p = 0.0002 (***). F, in situ hybridization for Sox10 and Foxd3 (both probes were mixed) after electroporation of scrambled siRNA (n = 33; from 7 independent experiments), siRNA-MMP14 (n = 56; from 11 independent experiments) or siRNA-MMP14 in combination with either MMP14-FL (n = 18; from 6 independent experiments), ΔCAT (n = 14; from 3 independent experiments), EA (n = 14; from 2 independent experiments) or ΔCyto (n = 14; from 3 independent experiments). Each electroporated condition is associated with a corresponding transversal section at the level of rhombomere 6 (dotted line). G, Mean area occupied by Sox10-positive neural crest cells above the neural tube in the whole rhombencephalic regions (normalized to scrambled siRNA). ANOVA followed by multiple comparisons, ****, p < 0.0001, ***, p = 0.004. Asterisks indicate the electroporated side. Scale bars, B-C, 50 μm; F, whole mount 50 μm, sections 25 μm.

Article Snippet: The following antibodies were used: mouse anti-Tubulin (Sigma, T5201; 1:10000), rabbit anti-snail2 (Cell Signaling, C19G7, 1/1000), rabbit anti-Histone H3 (Abcam ab1791, 1/5000), anti-rabbit-HRP or anti-mouse-HRP 1:10000 (Millipore).

Techniques: Activity Assay, Migration, In Situ Hybridization, Electroporation

Journal: Differentiation; research in biological diversity

Article Title: Delamination of chick cephalic neural crest cells requires an MMP14-dependent downregulation of Cadherin-6B.

doi: 10.1016/j.diff.2025.100836

Figure Lengend Snippet: Fig. 3. MMP14 loss-of-function leads to the accumulation of neural crest cells expressing snail2, cadherin-6B, rhoB, and integrin-β3 in the dorsal neural tube. A-F, in situ hybridization for snail2 (A, nscrambled = 7; nsiMMP14 = 4), rhoB (B, nscrambled = 6; nsiMMP14 = 6), cadherin-6B (C, nscrambled = 3; nsiMMP14 = 4), ets-1 (D, nscrambled = 3; nsiMMP14 = 4), cadherin-7 (E, nscrambled = 6; nsiMMP14 = 6), integrin-β3 (F, nscrambled = 8; nsiMMP14 = 6) at 12 h post electroporation. Scale bar in whole mount 100 μm, in sections 25 μm. Sections are at the level of rhombomere 6. Inserts show the electroporated region for each embryo. Asterisks indicate the electroporated side. Bracket in A indicate an expansion of the snail2 territory on the siRNA-MMP14 electroporated side. Arrowheads in C, D and F indicate neural crest cells in the dorsal neural tube expressing the corresponding gene of each panel. G-H, ratios of RhoB (G; nscrambled = 5 embryos, 22 sections; nsiRNA-MMP14 = 6 embryos, 38 sections, two- tailed Mann-Whitney test, p = 0.0173) and integrin-b3 (H; nscrambled = 6 embryos, 30 sections; nsiRNA-MMP14 = 7 embryos, 24 sections, two-tailed Mann-Whitney test, p = 0.0012) areas in electroporated and non-electroporated sides of the dorsal neural tube, averaged per embryo and normalized to the mean ratio in the scrambled siRNA condition.

Article Snippet: The following antibodies were used: mouse anti-Tubulin (Sigma, T5201; 1:10000), rabbit anti-snail2 (Cell Signaling, C19G7, 1/1000), rabbit anti-Histone H3 (Abcam ab1791, 1/5000), anti-rabbit-HRP or anti-mouse-HRP 1:10000 (Millipore).

Techniques: Expressing, In Situ Hybridization, Electroporation, Two Tailed Test, MANN-WHITNEY

Journal: Differentiation; research in biological diversity

Article Title: Delamination of chick cephalic neural crest cells requires an MMP14-dependent downregulation of Cadherin-6B.

doi: 10.1016/j.diff.2025.100836

Figure Lengend Snippet: Fig. 7. Snail2 protein expression in control and MMP14 knockdown neural crest cells A-B, Immunostaining against Snail2 (magenta) in embryos electroporated with scrambled control siRNA (A, n = 12) or with siRNA-MMP14 (B, n = 29) at 6 h post-electroporation. Electroporated cells are GFP+ (green). C-D, Immunostaining against Snail2 (magenta) in embryos electroporated with scrambled control siRNA (C, n = 23) or with siRNA-MMP14 (D, n = 27) at 18 h post-electroporation. Electroporated cells are GFP+ (green). DAPI counterstaining for nuclei is in grey. E, Procedure to measure Snail2 intensity. F, Ratios of Snail2 raw levels in elec troporated cells over neighboring control cells. Arrows indicate the position of the GFP + cells. One-way ANOVA, uncorrected Fisher’s LSD, scrambled vs siRNA at 6 h p = 0.7258, scrambled vs siRNA at 18 h p = 0.9053, siRNA at 6 h vs siRNA at 18 h p = 0.3814. Scale bars 20 μm.

Article Snippet: The following antibodies were used: mouse anti-Tubulin (Sigma, T5201; 1:10000), rabbit anti-snail2 (Cell Signaling, C19G7, 1/1000), rabbit anti-Histone H3 (Abcam ab1791, 1/5000), anti-rabbit-HRP or anti-mouse-HRP 1:10000 (Millipore).

Techniques: Expressing, Control, Knockdown, Immunostaining, Electroporation