Journal: PLoS ONE

Article Title: Macrophage Migration Inhibitory Factor Is Involved in Ectopic Endometrial Tissue Growth and Peritoneal-Endometrial Tissue Interaction In Vivo : A Plausible Link to Endometriosis Development

doi: 10.1371/journal.pone.0110434

Figure Lengend Snippet: PCNA immunostainingwas carried out on endometrial implants from KO/KO mice (A), WT/WT mice (B) and mice treated with ISO-1 (C) or rhMIF (D). Insets show general histological views of endometrial implants. Black arrows show CD74 positive cells. Scale bar,10 µm.

Article Snippet: To detect CD74, MIF receptor, and proliferating cell nuclear antigen (PCNA), sections were incubated for 90 min at room temperature with a rabbit polyclonal anti-human CD74 (GeneTex, Irvine, Canada) [1∶1600 dilution in PBS/bovine serum albumin (BSA) 0.02%/Tween-20 0.05%] or a rabbit polyclonal anti-human PCNA (GeneTex, Irvine, Canada) [1∶800 dilution in PBS/bovine serum albumin (BSA) 0.02%/Tween-20 0.05%].

Techniques:

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Spatially restricted dental regeneration drives pufferfish beak development

doi: 10.1073/pnas.1702909114

Figure Lengend Snippet: Localization of a dental progenitor cell niche within the pufferfish dental lamina. (A–C) P. baileyi PCNA immunohistochemistry reveals high levels of cellular proliferation within the oral epithelium. As replacement teeth progress from late-initiation (A) to morphogenesis (C), the new tooth generation (R1) buds from the dental lamina (B). Successive rounds of replacement show that the dental generations stack on one another within an enameloid outer casing (black line) (C). (D) Sox2 immunohistochemical labeling during dental replacement initiation depicts high levels of Sox2 within both the developing taste buds (TB) and the dental progenitor site located within the labial oral epithelium (dental lamina) (black arrowhead). (E) Double immunofluorescence treatment for Sox2/PCNA in T. niphobles shows low levels of PCNA expression within the Sox2+ cells of the presumptive dental progenitor niche and cells within the aboral dental lamina exhibiting high levels of PCNA. The horizontal dashed line depicts image stitching of two adjacent images. White arrowhead marks region of overlapping PCNA/Sox2 expression. (F) BrdU pulse/chase experiments (0.2 mM) show the incorporation of BrdU into dividing cells after 6 wk of treatment, with high levels of incorporation noted in the distal dental lamina next to the base of the beak (white arrowhead). (G) After a further 8-wk chase, label-retaining cells were found in the most superficial dental lamina cells (open arrowhead) but not in the distal dental lamina (white arrowhead). Label-retaining cells found in the dental epithelium of the developing tooth are indicated by a white arrow. Images in F and G are composites of multiple images taken at high magnification and stitched together. (H) DiI labeling of the labial oral epithelium in P. suvattii highlighted this region as a presumptive source of dental progenitor cells. DiI was detected within the outer dental epithelium of the tooth (white arrowhead) 72 h post DiI treatment. (I) As summarized in a schematic representation, we observed a continuous field of Sox2+ cells between the labial taste bud and the dental progenitor site, with cells from the latter migrating and contributing to the new dental generations. Black arrows represent the direction of cell movement. (J) Sox2/ABC double immunohistochemical labeling on adult C. travancoricus highlights epithelial Sox2+/ABC− (a′, white filled arrow), Sox2+/ABC+ (b′, white arrow), and Sox2−/ABC+ (c′, white arrowhead) regions within the dental lamina. Coexpression of these markers marks the site of activation of putative dental progenitors within the oral epithelium. Dashed line across (J) depicts image stitching of two adjacent images. Images are orientated with labial to the left and oral to the top. The dotted line in all images depicts the boundary of the oral epithelium and the end of the dental lamina. DM, dental mesenchyme; ODE, outer dental epithelium; R1–3, replacement tooth generations; RT, regenerating tooth; TB, labial taste bud. (Scale bars: 25 µm in A–E; 20 µm in F, a′–F, c′; 50 µm in G and H; 15 µm in I.)

Article Snippet: Rabbit anti-Sox2/mouse anti-PCNA and rabbit anti-Sox2/mouse anti-active β-catenin (1:500) (05-665; Merck) double immunofluorescence was carried out as described by Martin et al. ( 17 ).

Techniques: Immunohistochemistry, Immunohistochemical staining, Labeling, Immunofluorescence, Expressing, Pulse Chase, Activation Assay

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Spatially restricted dental regeneration drives pufferfish beak development

doi: 10.1073/pnas.1702909114

Figure Lengend Snippet: Conserved odontogenic signaling regulates dental regeneration in pufferfish. (A–I) Expression of well-documented odontogenic markers belonging to Sox (sox2, A); canonical Wnt signaling (β-catenin, B; lef1 C); Pitx (pitx2, D); Shh (E), Notch (hes1, F; notch3 G); Bmp (bmp2, H); and Fgf (fgf3, I) gene families in T. niphobles embryos. The thin arrow marks the site of presumptive dental progenitors, with expression of pitx2 (D), lef1 (C), and sox2 (A) within this region. The thick arrow marks the distal end of the dental lamina. The filled arrowhead highlights an opening within the osteodentine beak casing through which new odontogenic cells bud from the dental lamina. β-cat (B), shh (E), hes1 (F), notch3 (G), bmp2 (H), and fgf3 (I) are all expressed within the epithelium of the latest developing teeth. (J) A diagrammatic illustration of odontogenetically similar structures in various polyphyodonts [pufferfish, alligator (7, 16), cichlid (5), and catshark (43)]. Four main developmental regions are highlighted: presumptive dental progenitors, progenitor cell activation marked by the coexpression of Sox and Wnt signals, dental epithelium differentiation marked by the up-regulation of various developmental genes at the distal tip of the dental lamina and the growth of a tooth bud, and dental morphogenesis. The dotted line depicts the boundary of the oral epithelium and the end of the dental lamina. All images were taken from 14-µm sagittal paraffin-embedded sections. A, B, E, F, H, and I are from T. niphobles embryos at 50 dpf. C, D, and G are from embryos at 32 dpf. R1-2, replacement tooth generations; S, suture; TB, labial taste bud. (Scale bars: 50 µm in B–D, F, and G; 35 µm in A, E, H, and I.)

Article Snippet: Rabbit anti-Sox2/mouse anti-PCNA and rabbit anti-Sox2/mouse anti-active β-catenin (1:500) (05-665; Merck) double immunofluorescence was carried out as described by Martin et al. ( 17 ).

Techniques: Expressing, Activation Assay

Journal: Oncology Letters

Article Title: TCF12 activates MAGT1 expression to regulate the malignant progression of pancreatic carcinoma cells

doi: 10.3892/ol.2021.13180

Figure Lengend Snippet: Overexpression of MAGT1 promotes the proliferation, migration and invasion of pancreatic carcinoma cells. (A) Protein and (B) mRNA expression levels of MAGT1 in the BxPC-3 cell line transfected with pcDNA3.1-MAGT1, and pcDNA3.1-NC plasmids using western blot analysis and reverse transcription-quantitative PCR, respectively. Untransfected cells were used as a control. (C) Cell Counting Kit-8 assay was used to analyze cell viability in cells transfected with pcDNA3.1-NC and pcDNA3.1-MAGT1 at different time points (24, 48 and 72 h). (D) Representative western blots and quantification showing the protein expression levels of Ki67, and PCNA in the BxPC-3 cells were increased in cells transfected with pcDNA3.1-MAGT1 compared with that in cells transfected with pcDNA3.1-NC. (E) The BxPC-3 cells transfected with pcDNA3.1-MAGT1 had increased cell migration and invasion compared with that in cells transfected with pcDNA3.1-NC. (F) Representative western blots and quantification showing the protein expression level of MMP2, and MMP9 was significantly increased in the BxPC-3 cells transfected with pcDNA3.1-MAGT1 compared with that in cells transfected with pcDNA3.1-NC. *P<0.05, **P<0.01, ***P<0.001 vs. pcDNA3.1-NC group. MAGT1, magnesium transporter 1; NC, negative control.

Article Snippet: The following antibodies were used: Rabbit anti-MAGT1 (1:1,000; cat. no. NBP1-69683; Novus Biologicals, LLC); rabbit anti-TCF12 (1:5,000; cat. no. A300-754A; Bethyl Laboratories, Inc.); mouse anti-Ki67 (1:1,000; cat. no. 350502; BioLegend, Inc.); rabbit anti-PCNA (1:1,500; cat. no. LS-B402-50; LifeSpan BioSciences, Inc.); rabbit anti-MMP2 (1:4,000; cat. no. ab92536; Abcam); rabbit anti-MMP9 (1:15,000; cat. no. ab76003; Abcam); rabbit anti-GAPDH (1:2,500; cat. no. ab9485; Abcam), HRP-conjugated goat anti-rabbit IgG H&L (1:20,000; cat. no. ab6721; Abcam) and HRP-conjugated goat anti-mouse IgG H&L (1:10,000; cat. no. ab6789; Abcam).

Techniques: Over Expression, Migration, Expressing, Transfection, Western Blot, Real-time Polymerase Chain Reaction, Cell Counting, Negative Control

Journal: Oncology Letters

Article Title: TCF12 activates MAGT1 expression to regulate the malignant progression of pancreatic carcinoma cells

doi: 10.3892/ol.2021.13180

Figure Lengend Snippet: Knockdown of MAGT1 expression inhibits the proliferation, migration and invasion of pancreatic carcinoma cells. (A) Protein and (B) mRNA expression level of MAGT1 in BxPC-3 cells transfected with shMAGT1-1, sh-MAGT1-2 or sh-NC was analyzed using western blot analysis and reverse transcription-quantitative PCR, respectively. Untransfected cells were used as the control. (C) Cell Counting kit-8 assay was used to analyze cell viability in cells transfected with sh-NC and sh-MAGT1-2 at different time points (24, 48 and 72 h). (D) Representative western blots and quantification showing the protein expression levels of Ki67. and PCNA in BxPC-3 cells were decreased in cells transfected with sh-MAGT1-2 compared with that in cells transfected with sh-NC. (E) The BxPC-3 cells transfected with sh-MAGT1-2 had decreased cell migration and invasion compared with that in cells transfected with sh-NC. (F) Representative western blots and quantification showing the protein expression level of MMP2, and MMP9 was significantly increased in the BxPC-3 cells transfected with sh-MAGT1-2 compared with that in cells transfected with sh-NC. *P<0.05, **P<0.01, ***P<0.001 vs. sh-NC group. MAGT1, magnesium transporter 1; NC, negative control; sh, short hairpin.

Article Snippet: The following antibodies were used: Rabbit anti-MAGT1 (1:1,000; cat. no. NBP1-69683; Novus Biologicals, LLC); rabbit anti-TCF12 (1:5,000; cat. no. A300-754A; Bethyl Laboratories, Inc.); mouse anti-Ki67 (1:1,000; cat. no. 350502; BioLegend, Inc.); rabbit anti-PCNA (1:1,500; cat. no. LS-B402-50; LifeSpan BioSciences, Inc.); rabbit anti-MMP2 (1:4,000; cat. no. ab92536; Abcam); rabbit anti-MMP9 (1:15,000; cat. no. ab76003; Abcam); rabbit anti-GAPDH (1:2,500; cat. no. ab9485; Abcam), HRP-conjugated goat anti-rabbit IgG H&L (1:20,000; cat. no. ab6721; Abcam) and HRP-conjugated goat anti-mouse IgG H&L (1:10,000; cat. no. ab6789; Abcam).

Techniques: Expressing, Migration, Transfection, Western Blot, Real-time Polymerase Chain Reaction, Cell Counting, Negative Control

Journal: Oncology Letters

Article Title: TCF12 activates MAGT1 expression to regulate the malignant progression of pancreatic carcinoma cells

doi: 10.3892/ol.2021.13180

Figure Lengend Snippet: TCF12 promotes the proliferation, migration and invasion of pancreatic carcinoma cells by regulating the expression of MAGT1. (A) Cell Counting Kit-8 assay was used to determine the cell viability in cells transfected with si-NC, si-TCF12-2, si-TCF12-2+pcDNA3.1-NC and si-TCF12-2+pcDNA3.1-MAGT1-2 at different time points (24, 48 and 72 h. (B) Protein and mRNA expression levels of Ki67, and PCNA in BxPC-3 cells were decreased in cells transfected with si-TCF12-2 compared with that in cells transfected with si-NC group. Overexpression of MAGT1 reversed the protein expression level of Ki67 and PCNA. GAPDH served as the loading control. (C) The BxPC-3 cells transfected with si-TCF12-2 had decreased migration and invasion compared with that in cells transfected with si-NC. Overexpression of MAGT1 reversed the decreased migration and invasion. (D) Protein and mRNA expression level of MMP2, and MMP9 was significantly decreased in the BxPC-3 cells transfected with si-TCF12-2 compared with that in cells transfected with si-NC. The expression level of MMP2 and MMP9 were partly increased when co-transfected with pcDNA3.1-MAGT1, and si-TCF12-2. **P<0.01, ***P<0.001 vs. si-NC group; # P<0.05, ### P<0.001 vs. si-TCF12-2 + pcDNA3.1-NC group. NC, negative control; TCF12, transcription factor 12; MMP2, matrix metallopeptidase 2; si, small interfering.

Article Snippet: The following antibodies were used: Rabbit anti-MAGT1 (1:1,000; cat. no. NBP1-69683; Novus Biologicals, LLC); rabbit anti-TCF12 (1:5,000; cat. no. A300-754A; Bethyl Laboratories, Inc.); mouse anti-Ki67 (1:1,000; cat. no. 350502; BioLegend, Inc.); rabbit anti-PCNA (1:1,500; cat. no. LS-B402-50; LifeSpan BioSciences, Inc.); rabbit anti-MMP2 (1:4,000; cat. no. ab92536; Abcam); rabbit anti-MMP9 (1:15,000; cat. no. ab76003; Abcam); rabbit anti-GAPDH (1:2,500; cat. no. ab9485; Abcam), HRP-conjugated goat anti-rabbit IgG H&L (1:20,000; cat. no. ab6721; Abcam) and HRP-conjugated goat anti-mouse IgG H&L (1:10,000; cat. no. ab6789; Abcam).

Techniques: Migration, Expressing, Cell Counting, Transfection, Over Expression, Negative Control