Journal: Frontiers in physiology

Article Title: An Msx2 - Sp6-Follistatin Pathway Operates During Late Stages of Tooth Development to Control Amelogenesis.

doi: 10.3389/fphys.2020.582610

Figure Lengend Snippet: FIGURE 1 | Screening of representative genes involved in amelogenesis: LS8 cells were transfected with pCMVtag2-Flag-Msx2 then cultured for 48 h. Total RNA was isolated from the cells and subjected to reverse transcription PCR analysis. Upregulated genes are shown in red while downregulated ones are shown in green. The expression of Sp6, Sp3, Sprouty 2, Connexin 43, Wnt3, Tgfb1 and Enam (enamelin), Laminin 5 alpha 3 (lama3), as well as Msx2 itself, is up-regulated in the Msx2 overexpressing cells. In contrast, the expression of Tbx1, Amel (amelogenin), Fst (Follistatin) is downregulated in the Msx2 overexpressing cells, while Ambn (ameloblastin) expression is partially diminished, almost not affected. Gapdh is the housekeeping gene. C, cells transfected with control vector only; Msx2, cells are transfected with pCMVtag2-Flag-Msx2.

Article Snippet: Frontiers in Physiology | www.frontiersin.org 2 October 2020 | Volume 11 | Article 582610 For loss of function of Msx2, commercially available small interfering RNA for Msx2 (Msx2-siRNA) was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, United States).

Techniques: Transfection, Cell Culture, Isolation, Reverse Transcription, Expressing, Control, Plasmid Preparation

Journal: Frontiers in physiology

Article Title: An Msx2 - Sp6-Follistatin Pathway Operates During Late Stages of Tooth Development to Control Amelogenesis.

doi: 10.3389/fphys.2020.582610

Figure Lengend Snippet: FIGURE 2 | The expression of Sp6 and Fst is modulated early in response to Msx2: LS8 cells were transfected with Msx2 over-expression plasmid for two time points, 4 and 16 h. Total RNA was isolated from the cells and subjected to qPCR analysis. (A) Sp6 and (B) Fst could be detected as early as 4h after transfection by real time qPCR. By 16 h, there was a significant increase in the expression of Sp6 (A) with a corresponding decrease of Fst expression (B). Gapdh is the normalizing gene. C, cells transfected with control vector only. The experiment was conducted three times in replicates of 3. **p ≤0.005.

Article Snippet: Frontiers in Physiology | www.frontiersin.org 2 October 2020 | Volume 11 | Article 582610 For loss of function of Msx2, commercially available small interfering RNA for Msx2 (Msx2-siRNA) was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, United States).

Techniques: Expressing, Transfection, Over Expression, Plasmid Preparation, Isolation, Control

Journal: Frontiers in physiology

Article Title: An Msx2 - Sp6-Follistatin Pathway Operates During Late Stages of Tooth Development to Control Amelogenesis.

doi: 10.3389/fphys.2020.582610

Figure Lengend Snippet: FIGURE 3 | Loss of function of Msx2 and Sp6 in LS8 ameloblast-derived cells: (A) Both LS8 and G5 cells were overexpressed with Msx2 over-expression plasmid. Representative RT-PCR showing Sp6 is upregulated whereas Fst is downregulated in both cell lines after Msx2 overexpression. Bottom panel shows densitometric quantification of the bands. (B) The knockdown of Msx2 with siRNA relative to scrambled control shows downregulation of Msx2 and Sp6 and upregulation of Fst, in 48 and 72 h. RT-PCR results were normalized to Gapdh that served as an internal control and expression levels were relative to scrambled controls. **P < 0.01. Bottom panel shows a graph describing the trend of genes’ expression. (C) Lentiviral (ShRNA) gene knockdown assay, where Msx2-shRNA and Sp6-shRNA, and non-target-shRNA infected LS8 cells further confirm the siRNA results. qPCR results were normalized to Gapdh. Bottom panel shows densitometric quantification of the bands. Experiments were done in triplicates. **p ≤0.005; ***p ≤0.0001.

Article Snippet: Frontiers in Physiology | www.frontiersin.org 2 October 2020 | Volume 11 | Article 582610 For loss of function of Msx2, commercially available small interfering RNA for Msx2 (Msx2-siRNA) was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, United States).

Techniques: Derivative Assay, Over Expression, Plasmid Preparation, Reverse Transcription Polymerase Chain Reaction, Knockdown, Control, Expressing, shRNA, Infection

Journal: Frontiers in physiology

Article Title: An Msx2 - Sp6-Follistatin Pathway Operates During Late Stages of Tooth Development to Control Amelogenesis.

doi: 10.3389/fphys.2020.582610

Figure Lengend Snippet: FIGURE 4 | Msx2 is essential for Sp6 gene expression during late stage tooth development: In situ hybridization analyses of transcripts in wild type (A,C) and Msx2 deficient (B,D) first lower molar teeth at postnatal day P1 (A,B) and postnatal day P3 (C,D). Expression of Sp6 is reduced (B) relative to wild type (A). In contrast, a dramatic reduction of Sp6 expression, is observed in Msx2 deficient ameloblasts (D) compared to wild type at P3 (C). sa, secretory ameloblasts. Scale: X400 (N = 4).

Article Snippet: Frontiers in Physiology | www.frontiersin.org 2 October 2020 | Volume 11 | Article 582610 For loss of function of Msx2, commercially available small interfering RNA for Msx2 (Msx2-siRNA) was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, United States).

Techniques: Gene Expression, In Situ Hybridization, Expressing

Journal: Frontiers in physiology

Article Title: An Msx2 - Sp6-Follistatin Pathway Operates During Late Stages of Tooth Development to Control Amelogenesis.

doi: 10.3389/fphys.2020.582610

Figure Lengend Snippet: FIGURE 5 | Predicted model of Msx2 and Sp6 function during amelogenesis: We show that the two transcription factors Msx2 and Sp6 regulate each other’s expression (red arrows) and that Msx2 binds directly to Sp6 promoter (thick red arrow), suggesting that Sp6 is a direct target of Msx2. In addition, we show that Msx2 alone, like Sp6 (thick red inhibition lines) inhibits follistatin expression. In coordination with Sp6 (thin red inhibition line) the inhibition of follistatin expression is less robust. Collectively, these results raise the possibility that the Sp6 and Msx2 transcription factors interact closely with each other and work in a concerted manner to form part of a network that controls ameloblast life cycle and amelogenesis, in a cell autonomous manner.

Article Snippet: Frontiers in Physiology | www.frontiersin.org 2 October 2020 | Volume 11 | Article 582610 For loss of function of Msx2, commercially available small interfering RNA for Msx2 (Msx2-siRNA) was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, United States).

Techniques: Expressing, Inhibition

Journal: Frontiers in Microbiology

Article Title: Growth Coordination Between Butyrate-Oxidizing Syntrophs and Hydrogenotrophic Methanogens

doi: 10.3389/fmicb.2021.742531

Figure Lengend Snippet: The production of CH 4 and H 2 from butyrate oxidation (A) by a mesophilic coculture consisting of Syntrophomonas wolfei (S.w.) and Methanococcus maripaludis (M.m.) and their growth synchronization (B,C) . Three levels of syntroph-to-methanogen cell ratios (i.e., 5:1, 3:2, and 1:5) were created at the beginning by inoculating different population sizes to fresh medium, which were designated as the high, middle, and low cell ratio treatments, respectively. The relative growth (B) of the individual populations was estimated as the fold change of cell number at any timepoint relative to that at the beginning. Note that the change in Syntrophomonas – Methanococcus cell ratio over time (C) is shown in normal scale. The error bars in (A,B) indicate the positive values of standard deviation of four replicates.

Article Snippet: Syntrophomonas wolfei G311 (DSM102351), S. lipocalidus TGB-C1 (DSM12680), and M. maripaludis S2 (DSM14266) were purchased from German culture collection DSMZ (Braunschweig, Germany).

Techniques: Standard Deviation

Journal: Microorganisms

Article Title: How Capsular Exopolysaccharides Affect Cell Surface Properties of Lactic Acid Bacteria

doi: 10.3390/microorganisms8121904

Figure Lengend Snippet: Microscopic images of the cell suspensions of S. thermophilus ST143+ ( a , b ), ST143− ( c , d ), DGCC7984 ( e , f ), and W. cibaria DSM14295 ( g , h ) stained with India ink for CPS visualization, before (left) and after (right) ultrasonic treatment for CPS removal. Magnification: 1000×, scale bar: 10 µm. Arrows indicate microbial cell chains.

Article Snippet: S. thermophilus ST143+ and ST143− were provided by Christian Hansen A/S (Hørsholm, Denmark), S. thermophilus DGCC7984 by Danisco Deutschland GmbH (Niebüll, Germany), and W. cibaria DSM14295 by Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (Braunschweig, Germany).

Techniques: Staining

Journal: Microorganisms

Article Title: How Capsular Exopolysaccharides Affect Cell Surface Properties of Lactic Acid Bacteria

doi: 10.3390/microorganisms8121904

Figure Lengend Snippet: Comparison of the cells of S. thermophilus ST143+ ( a , b ), ST143− ( c , d ), and W. cibaria DSM14295 ( e , f ) before (left) and after (right) ultrasonic treatment for CPS removal as demonstrated by SEM images. Z: Z-ring; S: septum; scale bar: 500 nm.

Article Snippet: S. thermophilus ST143+ and ST143− were provided by Christian Hansen A/S (Hørsholm, Denmark), S. thermophilus DGCC7984 by Danisco Deutschland GmbH (Niebüll, Germany), and W. cibaria DSM14295 by Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (Braunschweig, Germany).

Techniques: Comparison

Journal: Microorganisms

Article Title: How Capsular Exopolysaccharides Affect Cell Surface Properties of Lactic Acid Bacteria

doi: 10.3390/microorganisms8121904

Figure Lengend Snippet: Cell surface hydrophobicity (CSH) of cells from S. thermophilus ST143+, ST143−, DGCC7984, and W. cibaria DSM14295 before (violet) and after ultrasonic treatment for CPS removal (orange). Mean values within the same strain with different superscripts differed significantly (α = 0.05).

Article Snippet: S. thermophilus ST143+ and ST143− were provided by Christian Hansen A/S (Hørsholm, Denmark), S. thermophilus DGCC7984 by Danisco Deutschland GmbH (Niebüll, Germany), and W. cibaria DSM14295 by Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (Braunschweig, Germany).

Techniques: Cell Surface Hydrophobicity