Journal: Journal of translational medicine

Article Title: MiR-30c-5p loss-induced PELI1 accumulation regulates cell proliferation and migration via activating PI3K/AKT pathway in papillary thyroid carcinoma.

doi: 10.1186/s12967-021-03226-1

Figure Lengend Snippet: Fig. 3 PELI1 activates the PI3K-AKT signaling pathways. A KEGG enrichment analysis showed that genes co-expressed with PELI1 are enriched in varied of pathway. B The protein levels of p-AKT, AKT, Ki-67 and MMP2 were assessed by western blotting in PELI1-overexpression or Peli1-silencing PTC cells. C Western blotting analysis showing the levels of PELI1 and p-AKT in the mouse tumor samples. D Representative IHC staining pictures of Ki-67 and MMP2 from mouse tumor sections (left panel), and the quantification of Ki-67 and MMP2 expression (right panel) (n = 3). The scale bars represent 50 µm. E Colony formation assay of PELI1-overexpression PTC cells after PI3K/AKT inhibition (LY294002; left panel), and the quantification was shown (right panel; n = 3). F Transwell assay was performed to estimate the cell migration of PELI1-overexpressing PTC cells after PI3K/AKT inhibition, the representative images (× 200) of Transwell assays were displayed (left panel), and the quantification was shown (right panel; n = 3). G Inhibition efficiency of PI3K/AKT and protein levels of Ki-67 and MMP2 in PELI1-overexpressing PTC cells after LY294002 treatment were determined by western blotting. The numbers are presented as fold increase over the LV-NC group. *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: Briefly, PTC tissue sections were incubated with an antibody to PELI1 (12,053–1- AP, diluted 1: 200; Proteintech, Rosemont, IL); mice tumor sections were incubated with an antibody to Ki-67 (ab16667, diluted 1: 200; Abcam, Cambridge, MA) or MMP2 (10,373–2-ap, diluted 1: 200; Proteintech) overnight at 4 °C, followed incubating by HPR-conjugated secondary antibody.

Techniques: Protein-Protein interactions, Western Blot, Over Expression, Immunohistochemistry, Expressing, Colony Assay, Inhibition, Transwell Assay, Migration

Journal: Journal of translational medicine

Article Title: MiR-30c-5p loss-induced PELI1 accumulation regulates cell proliferation and migration via activating PI3K/AKT pathway in papillary thyroid carcinoma.

doi: 10.1186/s12967-021-03226-1

Figure Lengend Snippet: Fig. 5 MiR-30c-5p inhibits PTC cell proliferation and migration by downregulating PELI1. A The cell proliferation of PTC cells with miR-30c-5p mimic or its control transfection was detected by CCK8 assay (n = 5). B Colony formation assays of PTC cells transfected with miR-NC or miR-30c-5p, and the quantification of colony formation was shown (n = 3). C Representative images of Transwell assays of PTC cells transfected with miR-30c-5p, and the quantification was shown (n = 3). D Representative images of scratch wound healing assays of PTC cells transfected with miR-30c-5p. E Transfection efficiency of increased PELI1 overexpression (oe-PELI1) was determined by western blot. F Colony formation analysis showed that oe-PELI1 could partially reverse the miR-30c-5p-mediated proliferation inhibition of PTC cells (n = 3). G Transwell analysis showed that oe-PELI1 significantly alleviated miR-30c-5p-mediated migration inhibition of PTC cells (n = 3). H p-AKT, AKT, Ki-67 and MMP2 protein levels were determined by western blot in each group. *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: Briefly, PTC tissue sections were incubated with an antibody to PELI1 (12,053–1- AP, diluted 1: 200; Proteintech, Rosemont, IL); mice tumor sections were incubated with an antibody to Ki-67 (ab16667, diluted 1: 200; Abcam, Cambridge, MA) or MMP2 (10,373–2-ap, diluted 1: 200; Proteintech) overnight at 4 °C, followed incubating by HPR-conjugated secondary antibody.

Techniques: Migration, Control, Transfection, CCK-8 Assay, Over Expression, Western Blot, Inhibition

Journal: Journal of translational medicine

Article Title: MiR-30c-5p loss-induced PELI1 accumulation regulates cell proliferation and migration via activating PI3K/AKT pathway in papillary thyroid carcinoma.

doi: 10.1186/s12967-021-03226-1

Figure Lengend Snippet: Fig. 6 EVs derived from miR-30c-5p-modified hUCMSC (miR-30c-5p-EVs) downregulate PELI1 expression in PTC cells. A miR-30c-5p-EVs and NC-EVs were observed under a transmission electron microscope (Scale bars: 200 nm), and the size distributions of these EVs were detected using the Nanoparticle Tracking Analysis. B Western blot analysis of TSG101 and HSP70 expression in miR-30c-5p-EVs and NC-EVs. Ponceau S staining served as a loading control. C The relative miR-30c-5p levels in miR-30c-5p-EVs and NC-EVs were detected by real-time PCR (n = 3). D Fluorescence was evaluated using laser confocal microscopy (Scale bars: 20 μm). E PTC cells treated with miR-30c-5p-EVs showed a significantly increased expression of miR-30c-5p in comparison with cells added with NC-EVs (n = 3). F, G Expression of PELI1 in PTC cells was assessed by real-time PCR (F) and Western blot (G). H p-AKT, AKT, Ki-67 and MMP2 protein levels were determined by western blot in each group. *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: Briefly, PTC tissue sections were incubated with an antibody to PELI1 (12,053–1- AP, diluted 1: 200; Proteintech, Rosemont, IL); mice tumor sections were incubated with an antibody to Ki-67 (ab16667, diluted 1: 200; Abcam, Cambridge, MA) or MMP2 (10,373–2-ap, diluted 1: 200; Proteintech) overnight at 4 °C, followed incubating by HPR-conjugated secondary antibody.

Techniques: Derivative Assay, Modification, Expressing, Transmission Assay, Microscopy, Western Blot, Staining, Control, Real-time Polymerase Chain Reaction, Fluorescence, Confocal Microscopy, Comparison

Journal: Journal of translational medicine

Article Title: MiR-30c-5p loss-induced PELI1 accumulation regulates cell proliferation and migration via activating PI3K/AKT pathway in papillary thyroid carcinoma.

doi: 10.1186/s12967-021-03226-1

Figure Lengend Snippet: Fig. 8 Proposed model of miR-30c-5p-EVs as new avenues for the treatment of PTC cancer. PELI1 was highly expressed in PTC cancer, while miR-30c-5p was poorly expressed. MiR-30c-5p could inhibit PTC cell proliferation and migration by negatively mediating the expression of the PELI1. MiR-30c-5p-EVs could significantly downregulate PELI1 expression and suppress the progression of PTC in vitro and in vivo, concomitant with reduced p-AKT, Ki-67 and MMP-2 expression

Article Snippet: Briefly, PTC tissue sections were incubated with an antibody to PELI1 (12,053–1- AP, diluted 1: 200; Proteintech, Rosemont, IL); mice tumor sections were incubated with an antibody to Ki-67 (ab16667, diluted 1: 200; Abcam, Cambridge, MA) or MMP2 (10,373–2-ap, diluted 1: 200; Proteintech) overnight at 4 °C, followed incubating by HPR-conjugated secondary antibody.

Techniques: Migration, Expressing, In Vitro, In Vivo

Journal: International Journal of Molecular Medicine

Article Title: The DR1-CSE/H 2 S system inhibits renal fibrosis by downregulating the ERK1/2 signaling pathway in diabetic mice

doi: 10.3892/ijmm.2021.5062

Figure Lengend Snippet: Activation of the DR1-CSE/H 2 S pathway inhibits HG-induced collagen deposition in MCs. The expression of COL1, α-SMA and MMP9 in the HG-induced MCs was determined by western blotting. The experiments were repeated at least three times. The results were expressed as the mean ± standard error of the mean. * P<0.05, ** P<0.01 vs. control group; # P<0.05, ## P<0.01 vs. HG group; & P<0.05, && P<0.01 vs. HG + SKF38393 group. DR1, dopamine 1 receptors; CSE, cystathionine-γ-lyase; HG, high glucose; MC, mesangial cell; COL1, collagen 1; α-SMA, α-smooth muscle actin.

Article Snippet: The primary antibodies for anti-CSE, cyclin D1, PCNA, P21, COL1, α-smooth muscle actin (α-SMA), and MMP9 were from ProteinTech Group, Inc. DR1 antibody was purchased from GeneTex, Inc.

Techniques: Activation Assay, Expressing, Western Blot, Control

Journal: American Journal of Cancer Research

Article Title: Overexpression of MEF2D contributes to oncogenic malignancy and chemotherapeutic resistance in ovarian carcinoma

doi:

Figure Lengend Snippet: Effect of MEF2D knockdown on invasion and migration of OC cells in vitro. A. The migration abilities of SKOV3 and OVCAR3 were measured through testing the wound closure after MEF2D knockdown using wound healing assays. B. Transwell assays were used to detect the migration and invasion abilities after MEF2D knockdown in SKOV3 and OVCAR3 cells. Original magnifications, × 200 and × 400. C. Effects of MEF2D knockdown on migration associated protein MMP9 were analyzed by western blotting in SKOV3 and OVCAR3 cells. Error bars represent the s.d. of triplicate measurements. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: The primary antibodies used in our study included anti-MEF2D, anti-cleaved caspase3 (Abcam, Cambridge, UK), anti-cyclinD1, anti-c-Myc, anti-caspase3 (Proteintech), anti-MMP9, anti-HPSE, anti-IKBKE (abclonal, Wuhan, China).

Techniques: Migration, In Vitro, Western Blot