Journal: Experimental biology and medicine (Maywood, N.J.)

Article Title: Nucleated red blood cells participate in myocardial regeneration in the toad Bufo Gargarizan Gargarizan .

doi: 10.1177/15353702211013297

Figure Lengend Snippet: Figure 8. Biochemical assay results. At indicated days after cautery-injury (dac), samples of blood and/or the apical tissue were collected to analyze relevant substances by enzyme-linked immunosorbent assay or Western blotting. Data are presented as mean SD, n ¼ 3, respectively; *P < 0.05 and **P < 0.01 vs. Ctrl group were calculated by Dunnett’s post hoc test after ANOVA. (a). Apical TNF-a. (b). Apical IL1b. (c). Apical IL6. (d). Apical IL11. (e). Serum IL11. (f). Serum cardiotrophin-1 (CT-1). (g). Apical CT-1. (h). Serum erythropoietin (EPO). (i). Apical vascular endothelial growth factor (VEGF). (j). Serum VEGF. (k). Apical matrix metalloproteinase 2 (MMP2). (l). Apical MMP9. ADU: arbitrary densitometric unit relative to glyceraldehyde-3-phosphate dehydrogenase (GAPDH).

Article Snippet: The membrane was blocked in Odyssey Blocking Buffer (LI-COR Biosciences, Lincoln, NE, USA), then incubated overnight at 4 C in rabbit antihuman matrix metalloproteinase 2 (MMP2) Ab diluent (1:1000; cat. no. 4022, Cell Signaling Technology, Danvers, MA, USA) or in rabbit anti-human MMP9 Ab diluent (1:1000; cat. no. 2270, Cell Signaling Technology), and finally incubated in IR Dye 800CW-goat anti-rabbit IgG (1:10,000; cat. no. 926–32211, LI-COR Biosciences).

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot

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: Applied biochemistry and biotechnology

Article Title: LncRNA LINC00665 Promotes Ovarian Cancer Cell Proliferation and Inhibits Apoptosis via Targeting miR-181a-5p/FHDC.

doi: 10.1007/s12010-022-03943-3

Figure Lengend Snippet: Fig. 2 LINC00665 knockdown inhibits migration and invasion of SKOV-3 and OVCAR-3 cells. Wound healing assay (A) and transwell assay (B) were conducted to assess the migration and invasion of SKOV-3 and OVCAR-3 cells transfected with sh-LINC00665 or sh-NC. (C) Western blot analysis was used to assess the expression of MMP-2 and MMP-9 proteins. Three independent experiments were conducted. The graphs showed the mean ± SD of at least three experiments. * P < 0.05, **P < 0.01

Article Snippet: Next, the membranes were blocked for 1 h with TBST containing 1% skimmed milk powder and then incubated overnight at 4 °C with rabbit antibodies against human FHDC1 (1:1000; cat. 1 3 no. NBP1-93,579; Novus Biologicals), MMP2 (1:1000; cat. no. 40994S; Cell Signaling Technology, Danvers, MA, USA), MMP9 (1:1000; cat. no. 13667S; Cell Signaling Technology), Cyclin D1 (1:1000; cat. no. 55506S; Cell Signaling Technology), p21 (1:1000; cat. no. 2947S; Cell Signaling Technology), Bcl-2 (1:1000; cat. no. ab185002; Abcam), Bax (1:500; cat. no. ab53154; Abcam), Cleaved Caspase-3 (1:500; cat. no. ab49822; Abcam), Cleaved Caspase-9 (1:1000; cat. no. 20750S; Cell Signaling Technology), or GAPDH (1:1000; cat. no. 5174S; Cell Signaling Technology).

Techniques: Knockdown, Migration, Wound Healing Assay, Transwell Assay, Transfection, Western Blot, Expressing

Journal: Nature cell biology

Article Title: Reprogramming of the tumour microenvironment by stromal PTEN-regulated miR-320.

doi: 10.1038/ncb2396

Figure Lengend Snippet: Figure 4 miR-320 regulates the secretome of MMFs. (a) Pten+/+ and Pten−/−MMFs were left untreated (lanes 1 and 2) or were transiently transfected with miR negative control (NC), miR-320 precursors (320) or anti-miR-320 precursor (anti-320), respectively. Conditioned media were examined by Western blotting with antibodies against the indicated proteins; thrombospondin 2 (THBS2) serves as an internal control. (b) Representative images of DB7 mammospheres in the presence of conditioned medium from Pten−/−MMFs expressing siRNA negative control (NC) (n = 4) or siRNA against Mmp9 (n = 4). MMP9 downregulation was verified by Western blot using MMP9 antibody. MMP2 was used as a loading control. Scale bars, 200 µm. The insets in all panels are magnified ×2.5. Quantification of migratory zones (indicated by arrows) expressed as mean area of migration ±s.d., ∗P < 0.01 (bar graphs). (c) Representative images of DB7 mammospheres in the presence of conditioned medium from Pten−/−MMFs precleared with IgG (n = 4) or IgG precoupled with α-MMP9 (n = 4). Western blots using MMP9

Article Snippet: Antibodies used were as follows: LOXL2, MMP2, EMILIN2 (1:500, Santa Cruz Biotechnology), THBS1, THBS2, SFRP1 (1:1,000, R&D Systems), ETS2 (1:1,000, rabbit polyclonal affinity purified), MMP9 (1:1,000, Abcam), BMP1 (1:500, Oncogene), β-actin, α-tubulin (1:10,000, Sigma Aldrich), CTSB, PTEN, AKT, P-AKTS473, JNK, P-JNKT183/Y185, ERK and P-ERKT202/Y204 (1:1,000, Cell Signaling).

Techniques: Transfection, Negative Control, Western Blot, Control, Expressing, Migration