Journal: International journal of oncology

Article Title: Synergistic inhibitory effects on hepatocellular carcinoma with recombinant human adenovirus Aspp2 and oxaliplatin via p53-independent pathway in vitro and in vivo.

doi: 10.3892/ijo.2017.4105

Figure Lengend Snippet: Figure 5. Synergistic influence of Aspp2-ad and oxaliplatin on morphology and CD31, PCNA protein expression in hepatocellular carcinoma xenograft models. (A) H&E staining. Red arrow, mitotic nucleus; black arrow, necrosis. (B) MI of different groups. (C) PCNA expression of tumors in different groups. (D) CD31 expression of tumors in different groups. The magnification of A, C and D was x200. Compared with control group, *P<0.05.

Article Snippet: Human Aspp2 recombinant adenovirus (Aspp2-ad) was constructed by the Beijing Institute of Hepatology (Beijing, China); oxaliplatin was obtained from Qilu Pharmaceutical Co., Ltd. (Jinan, China); trypsin-ethylenediaminetetraacetic acid and Dulbecco's modified Eagle's medium (DMEM) were purchased from Gibco (Grand Island, NY, USA); fetal bovine serum (FBS) was from China Hangzhou Sijiqing Biological Technology Co., Ltd. (Hangzhou, China); 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) and dimethyl sulfoxide (DMSO) were provided by Sigma-Aldrich (St. Louis, MO, USA); p53 primary monoclonal antibody was obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA); activation of caspase-3, Bax, Bcl-2, LC3B, Beclin1, ERK, p-ERK, mTOR, p-mTOR, STAT3, p-STAT3 and GAPDH primary monoclonal antibody were from Cell Signaling Technology (Boston, MA, USA); CD31 and PCNA immuno-histochemistry kits were obtained from Beijing Zhongshan Gold Bridge Technology Co., Ltd. (Beijing, China); Annexin V apoptosis Kit-PE kit was from SouthernBiotech (Birmingham, AL, USA).

Techniques: Expressing, Staining, Control

Journal: PLoS ONE

Article Title: Impact of Glutathione Peroxidase-1 Deficiency on Macrophage Foam Cell Formation and Proliferation: Implications for Atherogenesis

doi: 10.1371/journal.pone.0072063

Figure Lengend Snippet: Macrophages were collected after differentiation of thioglycollate-elicited mouse peritoneal macrophages for 3 days in 10 ng/ml MCSF. A left panel , macrophages (2,5×10 4 cells) were incubated with MCSF or oxLDL, respectively, and BrdU for another 48 hours. Subsequently, the proliferative activity was investigated with a BrdU-based chemiluminescence assay and expressed as relative proliferation rate relating to control cells from ApoE −/− mice without stimulus. Data represent means ± SD of 4 to 5 separate experiments. *p<0.05 or **p<0,01 above the histogram indicate statistically significant differences between the different genotypes and below the histogram compared with cells without treatment of MCSF or oxLDL. A right panel , macrophages from GPx-1 −/− ApoE −/− mice were pretreated with 10 µM ebselen for 1 h and incubated with 10 µg/ml oxLDL and BrdU for another 48 h. The proliferation rate was expressed as the relative proliferation rate relating to the proliferative activity of untreated cells. Data represent means ± SD of 3 independent experiments. B, macrophages of GPx-1 −/− ApoE −/− (upper panel) and ApoE −/− (lower panel) mice were pre-incubated with 75 µM PD98059 or 15 µM U0126, respectively, for 1 h and then incubated with 10 ng/ml MCSF (left) or 10 µg/ml oxLDL (right) and BrdU for another 48 h. The proliferation rate was expressed as the relative proliferation rate relating to the proliferative activity of untreated cells. Data represent means ± SD of 3 to 5 independent experiments. C, Representative immunohistochemical staining for the proliferation marker PCNA in atherosclerotic lesions of the aortic sinus demonstrating more pronounced positive nuclear staining in GPx-1 −/− ApoE −/− mice (left panel) than ApoE −/− mice (right panel). The vessel lumen is to the upper left-hand corner. The demarcation between intima and media is indicated by arrowheads.

Article Snippet: Polyclonal goat anti-apolipoprotein B antibody, monoclonal rat anti-F4/80 (clone CI:A3-1) antibody, polyclonal rabbit antibody to PCNA (proliferating cell nuclear antigen), polyclonal rabbit antibody to phospho-MEK1/2 (MAP2K1/2 pSer217/221), polyclonal rabbit antibody to phospho-ERK1/2 (p44/42 MAPK pThr202) and polyclonal rabbit antibody to phospho-p90RSK1 (RPS6KA1 pThr348) for immunohistochemistry were purchased from Acris Antibodies GmbH (Herford, Germany).

Techniques: Incubation, Activity Assay, Chemiluminescence Immunoassay, Control, Immunohistochemical staining, Staining, Marker

Journal: Journal of Ophthalmology

Article Title: Topical Vitamin C Promotes the Recovery of Corneal Alkali Burns in Mice

doi: 10.1155/2021/2406646

Figure Lengend Snippet: Vitamin C promotes the stemness of corneal epithelial stem cells and increases the CK3 expression in vivo . (a) Representative images of hematoxylin and eosin staining of injured corneas. (b) Immunohistochemical analyses of p63 expression. (c) The number of p63-positive cells in both groups. (d) Immunohistochemical analyses of proliferating cell nuclear antigen (PCNA) expression. (e) The number of PCNA-positive cells in both groups. (f) Immunohistochemical analyses of CK3 expression. (g) The IOD/area of CK3 in the cornea. IOD refers to the integral optical density. Abbreviations: Ctrl, control group (sterile water group); Vc, vitamin C group; ns, no significant difference between the indicated groups. ∗ means the comparison of Vc group and control group. ## and ### means the comparison with day 1. ## in G means the comparison with the Vc group on day 4. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ## p < 0.01, and ### p < 0.001. Scale bar = 50 μ m.

Article Snippet: The following primary antibodies were used for the immunohistochemical analyses: p63 (1 : 1,200, AB124762; Abcam, Cambridge, UK), CK3 (1 : 100, ab77869; Abcam), proliferating cell nuclear antigen (PCNA) (1 : 2,000, GB13010-1; Servicebio, Wuhan, China), alpha-smooth muscle actin ( α -SMA) (1 : 500, GB13044; Servicebio), CD31 (1 : 100; Sangon Biotech, Shanghai, China), and myeloperoxidase (MPO) (1 : 500, GB11224; Servicebio).

Techniques: Expressing, In Vivo, Staining, Immunohistochemical staining

Journal: Journal of the Endocrine Society

Article Title: The Orphan Nuclear Receptor Liver Homolog Receptor-1 (Nr5a2) Regulates Ovarian Granulosa Cell Proliferation

doi: 10.1210/js.2017-00329

Figure Lengend Snippet: Proliferation of mouse granulosa cells is compromised in Nr5a2-depleted follicles. (A, left) Amhr2 Cre depletion of Nr5a2 transcripts in granulosa cells from mouse antral follicles (n = 5). (A, middle and right) BrdU and PCNA expression in CON and cKO granulosa cells at 44 h after gonadotropin treatment, as evaluated by quantitative image analysis. Data are means ± standard error (SE); * P < 0.05, *** P < 0.001. (B) Representative images showing the expression of BrdU in CON and cKO ovaries at 44 hours after gonadotropin stimulation in vivo (n = 10/n = 6). (C) Representative images for PCNA at 44 hours after gonadotropin stimulation (n = 4). In this and subsequent figures, CON signifies control animals, cKO the granulosa-specific mouse knockout line (genotype Nr5a2f/fAmhr2 Cre/+ ). Original scale bars, 50 µM.

Article Snippet: We then treated with a rabbit polyclonal antibody against PCNA (SA194; Biomol, Hamburg, Germany): first antibody 1:200 in BSA 5%/PBS at 4°C overnight and with cyanine-3 (Cy3)-conjugated goat anti-rabbit IgG second antibody (111-165-144; Jackson ImmunoResearch Laboratories), diluted 1:400 in PBS 1× for 1 hour at room temperature.

Techniques: Expressing, In Vivo, Control, Knock-Out

Journal: Journal of the Endocrine Society

Article Title: The Orphan Nuclear Receptor Liver Homolog Receptor-1 (Nr5a2) Regulates Ovarian Granulosa Cell Proliferation

doi: 10.1210/js.2017-00329

Figure Lengend Snippet: FSHr is elevated in cKO granulosa cells, and exogenous estrogen cannot rescue proliferation relative it CON animals. (A, left) Abundance of FSHr transcripts in CON and cKO mice following gonadotropin stimulation (n = 5). (A, middle) The number of proliferating cells following 1 mg estrogen administration in vivo , as determined by quantitative imaging of BrdU expression. (A, right) The quantitative analysis of PCNA expression between granulosa cells from CON and cKO animals. Data are expressed as means ± SE of n = 4 animals/group. * P < 0.05. (B) Representative photomicrographs of BrdU incorporation in ovaries from estrogen-treated CON and cKO mice. (C) Representative photomicrographs showing PCNA expression in the ovaries of estrogen-treated CON and cKO mice. Original scale bars, 50 µM.

Article Snippet: We then treated with a rabbit polyclonal antibody against PCNA (SA194; Biomol, Hamburg, Germany): first antibody 1:200 in BSA 5%/PBS at 4°C overnight and with cyanine-3 (Cy3)-conjugated goat anti-rabbit IgG second antibody (111-165-144; Jackson ImmunoResearch Laboratories), diluted 1:400 in PBS 1× for 1 hour at room temperature.

Techniques: In Vivo, Imaging, Expressing, BrdU Incorporation Assay

Journal: Molecular Medicine Reports

Article Title: Long non-coding RNA Unigene56159 promotes glioblastoma multiforme cell proliferation and invasion through negatively regulating microRNA-194-5p

doi: 10.3892/mmr.2019.10852

Figure Lengend Snippet: Expression of proliferation- and invasion-related markers in GBM cells. (A) Correlation between long non-coding RNA Unigene56159 and PCNA, MMP-2 and MMP-9 expression in TCGA GBM database. (B) Following transfection with si-Unigene56159 vs. si-NC group, the protein and (C) mRNA expression levels of proliferation (PCNA) and invasion (MMP-2, MMP-9) related biomarkers were explored. *P<0.05 vs. si-NC group. (D) Immunofluorescent staining assay displayed a decreased level of MMP-2 in si-Unigene56159 transfected cells (magnification ×400). The experiment was repeated three times. GBM, glioblastoma multiforme; MMP, matrix metalloproteinase; NC, negative control; PCNA, proliferating cell nuclear antigen; siRNA, small interfering RNA; TCGA, The Cancer Genome Atlas.

Article Snippet: The membranes were incubated overnight at 4°C with the following primary antibodies: rabbit anti-proliferating cell nuclear antigen PCNA (Rabbit polyclonal antibody, cat. no. 10205-2-AP, 1:500; Wuhan Sanying Biotechnology), rabbit MMP-2 (Rabbit polyclonal antibody, cat. no 10373-2-AP, 1:500; Wuhan Sanying Biotechnology), rabbit anti-MMP-9 (Rabbit polyclonal antibody, cat. no 10375-2-AP, 1:500; Wuhan Sanying Biotechnology) and mouse anti-GAPDH (Mouse Monoclonal Antibody, cat. no. sc-47724, 1:1,000; Santa Cruz Biotechnology, Inc.), as the loading control.

Techniques: Expressing, Transfection, Staining, Negative Control, Small Interfering RNA

Journal: Antioxidants

Article Title: Oxidative Stress, NF-κB-Mediated Inflammation and Apoptosis in the Testes of Streptozotocin–Induced Diabetic Rats: Combined Protective Effects of Malaysian Propolis and Metformin

doi: 10.3390/antiox8100465

Figure Lengend Snippet: Expression of NF-κB (p65) protein ( a , b ) and mRNA ( c ); and the expression of TNF-α protein ( d , e ) and mRNA ( f ) in the testes of ( a ) NC, ( b ) DC, ( c ) D+MP, ( d ) D+Met and ( e ) D+MP+Met groups. For each biological replicate, a negative control was included, in which phosphate buffered saline was substituted for the primary antibody. A representative photomicrograph of the negative control is shown as ( f ) (magnification = ×100, scale bar = 200 µm). For quantitative data, values are mean ± SD, n = 8 (for staining intensity) and n = 6 (for mRNA expression). a p < 0.05 versus NC, b p < 0.05 versus DC, c p < 0.05 versus D+MP, d p < 0.05 versus D+Met (one-way ANOVA followed by Tukey’s post-hoc test). NC: normoglycaemic control, DC: diabetic control, D+MP: diabetic rats treated with Malaysian propolis, D+Met: diabetic rats treated with metformin, D+MP+Met: diabetic rats treated with Malaysian propolis + metformin, NF-κB: nuclear factor kappa B, TNF-α: tumour necrosis factor alpha.

Article Snippet: Testes sections were incubated with polyclonal primary antibodies for NF-kB(p65) (1:80), IL-1β (1:80), IL-10 (1:40), caspase-3 (1:150) and PCNA (1:50) (Cloud-Clone Corp, Houston, TX, USA) overnight at 4 °C, while incubation with TNF-α (1:120) polyclonal primary antibody was performed for 1 h at room temperature.

Techniques: Expressing, Negative Control, Staining