Journal: bioRxiv

Article Title: Microsecond pulse electrical stimulation modulates cell migration

doi: 10.1101/2022.10.23.513372

Figure Lengend Snippet: (a) A graphical illustration showed the effects of μsPEF (pulse width:20 μs, frequency: 10 Hz, duration: 5 s) on the skin wound, promoting cell migration and extracellular matrix remodeling. (b) Representative time-lapse images showing the fibroblasts morphology after μsPEF (i.e., 750 and 1500 V/cm) treatment of different intensity at different time points (i.e., 0, 1 and 2 h). Detached cells are marked by yellow arrow. Scale bar, 50 μm. (c) The line graph representing the cell migration average speed per 1 h from 0 to 12 h after different intensity μsPEF (i.e., 750 and 1500 V/cm) treatment (blue circle: control; orange square: 750 V/cm; pink triangle: 1500 V/cm). Results are presented as mean ± standard deviation with 95% CI (n CTRL =42 cells, n 750 v/cm =48 cells, n 1500 v/cm =47 cells); * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001 versus control by one-way ANOVA for multiple comparisons. (d) Box plot showing the average cell migration speed with different intensity μsPEF (i.e., 750 and 1500 V/cm) and control treatments in 24 h. Results are presented as mean ± standard deviation with 95% CI (n CTRL =470 cells, n 750 V/cm =979 cells, n 1500 V/cm =535 cells). (e) Effects of μsPEF on the secretion of COLA2. The level of collagen type I α2 in cellular supernatants was measured after 48 h. The concentration of COLA2 was 0.109 ± 0.018 ng/mL in the control group, 0.257 ± 0.058 ng/mL in the 750 V/cm group and 0.363 ± 0.034 ng/mL in the 1500 V/cm group. Results are presented as mean ± standard deviation with 95% CI (n=5); * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001 versus control by one-way ANOVA for multiple comparisons. (f) Effects of μsPEF on the expression of FGF2. The level of FGF2 in cellular supernatants was measured after 48 h. The concentration of FGF2 was 44.139 ± 0.360 ng/mL in the control group, 48.012 ± 1.488 ng/mL in the 750 V/cm group and 48.523 ± 1.944 ng/mL in the 1500 V/cm group. Results are presented as mean ± standard deviation with 95% CI (n=3); ns=0.7329, * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001 versus control by one-way ANOVA for multiple comparisons.

Article Snippet: Then, the cells were cultured in serum-free medium for 48 h. The content of type I α collagen and basic fibroblast growth factor (FGF-2) in the supernatant were measured using commercially available Human COL1A2 (Collagen Type I Alpha 2) ELISA Kit (Elabscience, Wuhan, China) and Human bFGF/FGF2 (Basic Fibroblast Growth Factor) ELISA Kit (Elabscience, Wuhan, China) according to the manufacturer’s protocol respectively.

Techniques: Migration, Control, Standard Deviation, Concentration Assay, Expressing

Journal: Life

Article Title: The Inhibitory Effect of Noscapine on the In Vitro Cathepsin G-Induced Collagen Expression in Equine Endometrium

doi: 10.3390/life11101107

Figure Lengend Snippet: Levels of significance ( p values) for two- and three-way interactions between estrous cycle phases, treatment time, and cathepsin G (CAT) or noscapine (NOSC) treatments in the analyses of relative transcript COL1A2 gene and COL1 protein relative abundance. The results were considered significant at p < 0.05.

Article Snippet: The COL1 primary antibody (1:1000 diluted; RRID: AB_2891017, 20121, Novotec, Lyon, France) was incubated overnight at 4 °C and previously validated to equine endometrium by Rebordão et al. [ ].

Techniques: Isolation

Journal: Life

Article Title: The Inhibitory Effect of Noscapine on the In Vitro Cathepsin G-Induced Collagen Expression in Equine Endometrium

doi: 10.3390/life11101107

Figure Lengend Snippet: Inhibition of cathepsin G (0.1 or 1 µg/mL) by noscapine (NOSC; 45 µg/mL) on relative collagen type I alpha 2 chain ( COL1A2 ) mRNA transcription and collagen type I (COL1) protein relative abundance in mare endometrial explants, regardless of estrous cycle phase and time of treatment. Results were considered significant at p < 0.05 and are displayed as least square means ± SEM. Different superscript letters indicate significant differences between CAT concentrations (a, b: CAT 0.1 µg/mL ≠ CAT 1 µg/mL, p < 0.05; c, d: CAT 0.1 µg/mL + NOSC ≠ CAT 1 µg/mL + NOSC, p < 0.05). Asterisks alone represent significant differences relative to the respective control and asterisks above the connecting lines indicate significant differences between treatments (** p < 0.01; *** p < 0.001).

Article Snippet: The COL1 primary antibody (1:1000 diluted; RRID: AB_2891017, 20121, Novotec, Lyon, France) was incubated overnight at 4 °C and previously validated to equine endometrium by Rebordão et al. [ ].

Techniques: Inhibition, Control

Journal: Life

Article Title: The Inhibitory Effect of Noscapine on the In Vitro Cathepsin G-Induced Collagen Expression in Equine Endometrium

doi: 10.3390/life11101107

Figure Lengend Snippet: Effect of cathepsin G (CAT; 0.1 or 1 µg/mL), noscapine (NOSC; 45 µg/mL), or CAT (0.1 or 1 µg/mL) + NOSC (45 µg/mL) treatments in explants of mare endometrium from follicular phase (FP) or mid-luteal phase (MLP) for 24 or 48 h on relative collagen type I alpha 2 chain ( COL1A2 ) mRNA transcription ( A , B ) and collagen type I (COL1) protein relative abundance ( C , D ). Results were considered significant at p < 0.05 and shown as least square means ± SEM. Asterisks alone represent significant differences relative to the respective control and asterisks above connecting lines indicate significant differences of CAT + NOSC treatment relative to the respective CAT-treated group (* p < 0.05; ** p < 0.01; *** p < 0.001).

Article Snippet: The COL1 primary antibody (1:1000 diluted; RRID: AB_2891017, 20121, Novotec, Lyon, France) was incubated overnight at 4 °C and previously validated to equine endometrium by Rebordão et al. [ ].

Techniques: Control

Journal: Life

Article Title: The Inhibitory Effect of Noscapine on the In Vitro Cathepsin G-Induced Collagen Expression in Equine Endometrium

doi: 10.3390/life11101107

Figure Lengend Snippet: Effect of cathepsin G (CAT; 0.1 or 1 μg/mL), noscapine (NOSC; 45 μg/mL), or CAT (0.1 or 1 μg/mL) + NOSC (45 μg/mL) treatments on relative collagen type I alpha 2 chain ( COL1A2 ) mRNA transcription ( A ) and collagen type I (COL1) protein relative abundance ( B ) in equine endometrial explants treated for 24 or 48 h, regardless of estrous cycle phase. Results are shown as least square means ± SEM and considered significant at p < 0.05. Asterisks above connecting lines indicate significant differences of the same treatment between time of treatment or differences between different concentrations of CAT at the same time of treatment (* p < 0.05; ** p < 0.01).

Article Snippet: The COL1 primary antibody (1:1000 diluted; RRID: AB_2891017, 20121, Novotec, Lyon, France) was incubated overnight at 4 °C and previously validated to equine endometrium by Rebordão et al. [ ].

Techniques:

Journal: Life

Article Title: The Inhibitory Effect of Noscapine on the In Vitro Cathepsin G-Induced Collagen Expression in Equine Endometrium

doi: 10.3390/life11101107

Figure Lengend Snippet: Effect of cathepsin G (CAT; 0.1 or 1 μg/mL), noscapine (NOSC; 45 μg/mL), or CAT (0.1 or 1 μg/mL) + NOSC (45 μg/mL) treatments on relative collagen type I alpha 2 chain ( COL1A2 ) mRNA transcription ( A ) and collagen type I (COL1) protein relative abundance ( B ) in explants of mare endometrium from follicular (FP) or mid-luteal (MLP) phases, regardless of treatment time. Results are shown at least square means ± SEM and considered significant at p < 0.05. Asterisks above connecting lines indicate significant differences of the same treatment between estrous cycle phase or differences between different concentrations of CAT at the same estrous cycle phase (* p < 0.05; ** p < 0.01; *** p < 0.001).

Article Snippet: The COL1 primary antibody (1:1000 diluted; RRID: AB_2891017, 20121, Novotec, Lyon, France) was incubated overnight at 4 °C and previously validated to equine endometrium by Rebordão et al. [ ].

Techniques:

Journal: Stem Cell Research & Therapy

Article Title: Connective tissue growth factor promotes cementogenesis and cementum repair via Cx43/β-catenin axis

doi: 10.1186/s13287-022-03149-8

Figure Lengend Snippet: CTGF promoted cementogenesis of hPDLSCs in vitro. A ARS staining showed a greater increase in mineralized nodules in hPDLSCs treated with the ascending-concentration of CTGF relative to the control group on the 14th day. B ALP staining on day 7 demonstrated the enhanced ALP activity in hPDLSCs after CTGF treatment. C Representative WB results of the mineralization-related proteins showed increasing expression of Osx, Runx2, CAP and Col1 in hPDLSCs when stimulated with the concentration-ascending CTGF on the 14th day. D – G Representative charts indicating the quantification of Osx, Runx2, CAP and Col1 in Fig. C. n = 3. Data are obtained from three independent images. * p < 0.05. H – L qPCR data showed that the expression of mineralization-related factors (Runx2, Osx, CAP, CEMP, ALP) enhanced in hPDLSCs treated with the concentration-ascending CTGF on the 14th day. Data are obtained from three independent results. * p < 0.05

Article Snippet: After incubation with 5% BSA, the sections were incubated overnight with anti-Col1 (1:200, 501,352, Zen-Bio, China), anti-Osterix (1:200, Huabio, ER1914-47, China), anti-Runx2 (1:200, 860,139, Zen-Bio, China), anti-CAP (1:200, sc-53947, Santa, America), anti-Cx43 (1:200, 340,279, Zen-Bio, China), and anti-active β-catenin (1:200, 19,807, Cell Signaling Technology, America) antibodies, followed by incubation with the corresponding IgG secondary antibodies.

Techniques: In Vitro, Staining, Concentration Assay, Activity Assay, Expressing

Journal: Stem Cell Research & Therapy

Article Title: Connective tissue growth factor promotes cementogenesis and cementum repair via Cx43/β-catenin axis

doi: 10.1186/s13287-022-03149-8

Figure Lengend Snippet: Cx43 expression was consistent with the expression of mineralization-related proteins induced by CTGF in vivo. A , B Representative IHC staining illustrated the protein expression of Osx and Runx2 with or without the treatment of CTGF on 0th day, 14th and 28th day. Images derived from three independent data. C , D Representative IHC staining illustrated the protein expression of CAP and Col1 with or without the treatment of CTGF on 0th day, 14th and 28th day. Images derived from three independent data. E , F The means of IOD of Osx and Runx2 in Fig. A, B. * p < 0.05. G , H The means of IOD of CAP and Cx43in Fig. A, B. * p < 0.05. I IHC results represented that the protein abundance of Cx43 with or without the treatment of CTGF on 0th day, 14th and 28th day. Images derived from three independent data. J The means of IOD of CAP and Cx43in Fig. A, B. * p < 0.05. K IF results represented that the abundance of Cx43 in CTGF-treated periodontal tissues. Cx43, red; nucleus, blue

Article Snippet: After incubation with 5% BSA, the sections were incubated overnight with anti-Col1 (1:200, 501,352, Zen-Bio, China), anti-Osterix (1:200, Huabio, ER1914-47, China), anti-Runx2 (1:200, 860,139, Zen-Bio, China), anti-CAP (1:200, sc-53947, Santa, America), anti-Cx43 (1:200, 340,279, Zen-Bio, China), and anti-active β-catenin (1:200, 19,807, Cell Signaling Technology, America) antibodies, followed by incubation with the corresponding IgG secondary antibodies.

Techniques: Expressing, In Vivo, Immunohistochemistry, Derivative Assay

Journal: Stem Cell Research & Therapy

Article Title: Connective tissue growth factor promotes cementogenesis and cementum repair via Cx43/β-catenin axis

doi: 10.1186/s13287-022-03149-8

Figure Lengend Snippet: Positive cementogenesis induced by CTGF was reversed by ablation of Cx43. A Western blotting demonstrated the abundance of Cx43 in hPDLSCs treated with Si-Cx43. B Bar chart showed the levels of Cx43 in Fig. A ratio to the untreated groups. Data are obtained from three independent images. n = 3, * p < 0.05. C Representative images of ARS indicated the attenuated ability of mineralization induced by Si-Cx43 in hPDLSCs compared with the untreated groups. D Representative images of ARS staining showed the CTGF-mediated enhancement in mineralized nodules was reversed by the addition of Si-Cx43. E Representative images of ALP staining showed the CTGF-mediated enhancement in ALP activity was reversed by Si-Cx43. F Representative images of the data of mineralization-associated proteins (Runx2, Osx, CAP, Col1) and Cx43 obtained by western blotting showed that increased mineralization capacity of CTGF expression was reverted by Si-Cx43. G – K Bar chart showed the levels of Runx2, Osx, CAP, Col1 and Cx43 in Fig. F ratio to the untreated control. Data are obtained from three independent images. n = 3, * p < 0.05. L–P mRNA expression of Runx2, Osx, ALP, CAP and CEMP, as detected by PCR ( n = 3) as detected by PCR

Article Snippet: After incubation with 5% BSA, the sections were incubated overnight with anti-Col1 (1:200, 501,352, Zen-Bio, China), anti-Osterix (1:200, Huabio, ER1914-47, China), anti-Runx2 (1:200, 860,139, Zen-Bio, China), anti-CAP (1:200, sc-53947, Santa, America), anti-Cx43 (1:200, 340,279, Zen-Bio, China), and anti-active β-catenin (1:200, 19,807, Cell Signaling Technology, America) antibodies, followed by incubation with the corresponding IgG secondary antibodies.

Techniques: Western Blot, Staining, Activity Assay, Expressing

Journal: Stem Cell Research & Therapy

Article Title: Connective tissue growth factor promotes cementogenesis and cementum repair via Cx43/β-catenin axis

doi: 10.1186/s13287-022-03149-8

Figure Lengend Snippet: Negative cementogenesis induced by Si-CTGF in hPDLSCs was motivated by the Cx43 agonist. A Western blotting demonstrated the abundance of CTGF in hPDLSCs treated with Si-CTGF. B Bar chart showed the levels of CTGF in A ratio to the untreated groups. Data are obtained from three independent images. n = 3, * p < 0.05. C Western blotting demonstrated the abundance of Cx43 in hPDLSCs treated with Cx43 agonist ATRA. D Bar chart showed the levels of Cx43 in C ratio to the untreated groups. Data are obtained from three independent images. n = 3, * p < 0.05. E Representative images of ARS indicated the attenuated ability of mineralization induced by Si-CTGF in hPDLSCs was regained by Cx43 agonist ATRA when compared with the untreated groups. F Representative images of ALP staining showed the attenuated ALP activity stimulated by Si-CTGF in hPDLSCs was regained by Cx43 agonist ATRA when in comparison to the untreated groups. G Representative image of the data of mineralization-associated proteins (Runx2, Osx, CAP, Col1) and Cx43 obtained by western blotting showed that the decreased mineralization capacity of CTGF expression was reverted by Cx43 agonist ATRA. H , I Bar chart showed the protein levels of Runx2, Osx, CAP, Col1 and Cx43 in G ratio to the untreated groups. Data are obtained from 3 independent images. n = 3, * p < 0.05. J mRNA expression of Osx, Runx2, ALP, CAP and CEMP, as detected by PCR ( n = 3) as detected by PCR

Article Snippet: After incubation with 5% BSA, the sections were incubated overnight with anti-Col1 (1:200, 501,352, Zen-Bio, China), anti-Osterix (1:200, Huabio, ER1914-47, China), anti-Runx2 (1:200, 860,139, Zen-Bio, China), anti-CAP (1:200, sc-53947, Santa, America), anti-Cx43 (1:200, 340,279, Zen-Bio, China), and anti-active β-catenin (1:200, 19,807, Cell Signaling Technology, America) antibodies, followed by incubation with the corresponding IgG secondary antibodies.

Techniques: Western Blot, Staining, Activity Assay, Expressing