Journal: PLoS ONE

Article Title: Is prnt a Pseudogene? Identification of Ram Prt in Testis and Ejaculated Spermatozoa

doi: 10.1371/journal.pone.0042957

Figure Lengend Snippet: APPA labeling of ram germinal cells in seminiferous tubules (E). Positive cells were detected (arrows) in spermatogonia (C), spermatocytes (D), spermatides (A), and spermatozoa (B). Negative control using mouse control serum (pre-immune), as the primary antibody (F). Same results were obtained when incubating sections with the secondary antibody alone (data not shown). Vimentin (G) and Ki-67 antigen (H), immunodetection of Sertoli cells cytoskeletal and spermatides, respectively. Spermatogonia (Sg), spermatocytes (Sc), spermatides (Sp), spermatozoa (Spz), Sertoli cell (S). Magnifications (A, B, C and D) ×1000, (E and F) ×100.

Article Snippet: To precisely determine the different cellular types present at the seminiferous tubules, and thus facilitate the identification of the cells expressing Prt, two other antibodies were used: one directed against Vimentin (Monoclonal Mouse Anti-Vimentin, clone Vim 3B4, Code M7020, Dako, Dilution 1/200), and the other against Ki-67 (Monoclonal Mouse Anti-Human Ki-67 antigen, clone MIB-1, Code M7240, Dako, Dilution 1/100), to locate respectively the Sertoli cell cytoskeleton, and the germinal cells with particular emphasis to the spermatides, as Ki-67 is a nuclear protein that is mainly expressed in proliferating cells .

Techniques: Labeling, Negative Control, Immunodetection

Journal: BMC Urology

Article Title: Inhibition of COX-2 expression by topical diclofenac enhanced radiation sensitivity via enhancement of TRAIL in human prostate adenocarcinoma xenograft model

doi: 10.1186/1471-2490-13-1

Figure Lengend Snippet: Diclofenac induced the tumor growth delay and enhanced the effect of RT in xenograft mouse model of LNCaP-COX-2. The xenograft mice bearing LNCaP-COX-2 tumor were treated with topical administration of diclofenac gel onto mice skin with cotton swab once a day (day 1–36). The tumors in RT alone group and combination group were exposed to IR with a single dose of 3 Gy on day 3. Tumor volume was measured with calipers and calculated as 0.52LW 2 . ( A ) Tumor volume was significantly reduced in combination group. ( B ) Immunohistochemical staining for COX-2 and Ki-67 was conducted. The expression of COX-2 decreased in diclofenac group and combination group compared with control group, while the expression of COX-2 increased in RT group. The expression of Ki-67 significantly decreased in diclofenac group and combination group. Original magnification, x200.

Article Snippet: Next, the tissue sections were incubated with mouse monoclonal anti-human COX-2 antibody (Santa Cruz, USA) and mouse monoclonal anti-human Ki-67 antibody (Dako, Glostrup, Denmark).

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: Scientific Reports

Article Title: Combination of ruthenium(II)-arene complex [Ru(η 6 - p -cymene)Cl 2 (pta)] (RAPTA-C) and the epidermal growth factor receptor inhibitor erlotinib results in efficient angiostatic and antitumor activity

doi: 10.1038/srep43005

Figure Lengend Snippet: ( A ) Tumor growth curves of A2780 tumors grafted on the CAM showing tumor volume with respect to treatment day represented as the percentage of the final control tumor volume. S indicates synergy (CI < 1). N = 30 in the control group and N = 4–14 in the treatment groups (two-way ANOVA: F(6,555) = 8.994, P < 0.0001). ( B ) Representative images of resected tumors in each treatment group. Control tumors were treated with 0.14% DMSO in NaCl. ( C ) Tumor weight (mg) after resection on treatment day 8 (when the experiment was terminated (one-way ANOVA: F(2, 48) = 3.35, P = 0.04)). ( D ) Microvessel density (MVD) analysis measured as the number of vessels per mm 2 of vascularized tumor area and represented as percentage of control (one-way ANOVA: F(2, 25) = 15.23, P < 0.0001). ( E ) Quantification of the percentage of whole tumor sections that are positive for the proliferation marker Ki67 (one-way ANOVA: F(2, 27) = 4.25, P = 0.025) ( F ) Representative images of immunohistochemical (IHC) staining for the endothelial cell marker CD31 (brown) counter-stained with haematoxylin (purple/blue). ( G ) Representative images of IHC staining for the proliferation marker Ki67 (brown/orange) counter-stained with haematoxylin (purple/blue). I indicates the combination erlotinib 20 μg/kg/day + RAPTA-C 21.6 μg/kg/day and II indicates the combination erlotinib 10 μg/kg/day + RAPTA-C 216 μg/kg/day in all graphs. Error bars represent SEM and *P < 0.05, **P < 0.01 indicate significance versus CTRL. Statistical analysis was performed using a two-way ANOVA with post-hoc Tukey’s multiple comparison test ( A ) or one-way ANOVA with post-hoc Dunnett’s multiple comparison test between the combination and control ( C – E ).

Article Snippet: Sections were stained separately for proliferation marker with primary antibodies against Ki67 (rabbit anti-human Ki67, 1:100, clone SP6; Thermo Scientific) and subsequent incubation with secondary swine anti-rabbit biotinylated antibodies (1:200; Dako, Glostrup, Denmark).

Techniques: Control, Marker, Immunohistochemical staining, Immunohistochemistry, Staining, Comparison

Journal: Scientific Reports

Article Title: Combination of ruthenium(II)-arene complex [Ru(η 6 - p -cymene)Cl 2 (pta)] (RAPTA-C) and the epidermal growth factor receptor inhibitor erlotinib results in efficient angiostatic and antitumor activity

doi: 10.1038/srep43005

Figure Lengend Snippet: ( A ) Tumor growth curves of A2780cisR tumors grafted on the CAM showing tumor volume with respect to treatment day, represented as the percentage of the final control tumor volume. S indicates synergy. N = 20 in the control group and N = 7–15 in the treatment groups. Control tumors were treated with 0.14% DMSO in NaCl (two-way ANOVA: F(6,550) = 4.692, P = 0.0001). ( B ) Tumor weight (mg) after resection at treatment day 8 (when the experiment was terminated; not significant, one-way ANOVA: F(2,54) = 3.05, P = 0.055). ( C ) Microvessel density (MVD) analysis measured as the number of vessels per mm 2 of vascularized tumor area and represented as percentage of control (one-way ANOVA: F(2,31) = 5.641, P = 0.0081). ( D ) Quantification of the percentage of whole tumor surfaces that are positive for the proliferation marker Ki67 (one-way ANOVA: F(2,34) = 4.67, P = 0.016). ( E ) Representative images of IHC staining for the endothelial cell marker CD31 (brown) counter-stained with haematoxylin (purple/blue). ( F ) Representative images of IHC staining for the proliferation marker Ki67 (brown/orange) counter-stained with haematoxylin (purple/blue). I indicates the combination erlotinib 20 μg/kg/day + RAPTA-C 21.6 μg/kg/day and II indicates the combination 10 μg/kg/day + RAPTA-C 216 μg/kg/day in all graphs. Error bars represent SEM and *P < 0.05, **P < 0.01 indicate significance versus CTRL. Statistical analysis was performed using a two-way ANOVA with post-hoc Tukey’s multiple comparison test ( A ), or one-way ANOVA with post-hoc Dunnett’s multiple comparison test between the combination and control ( B – D ).

Article Snippet: Sections were stained separately for proliferation marker with primary antibodies against Ki67 (rabbit anti-human Ki67, 1:100, clone SP6; Thermo Scientific) and subsequent incubation with secondary swine anti-rabbit biotinylated antibodies (1:200; Dako, Glostrup, Denmark).

Techniques: Control, Marker, Immunohistochemistry, Staining, Comparison

Journal: Scientific Reports

Article Title: Combination of ruthenium(II)-arene complex [Ru(η 6 - p -cymene)Cl 2 (pta)] (RAPTA-C) and the epidermal growth factor receptor inhibitor erlotinib results in efficient angiostatic and antitumor activity

doi: 10.1038/srep43005

Figure Lengend Snippet: ( A ) Resected tumor volume of each treatment group as the percentage control on treatment day 12. N = 5–7 in all treatment groups. Control tumors were treated i.p. with 4% DMSO in NaCl (One-way ANOVA with Tukey’s multiple comparisons test: F(5,33) = 4.145, P = 0.0050). Pharmacokinetic studies in mice revealed a plasma half-life time of 1.5–3 hours after ingestion for erlotinib and 10.39–12.21 hours after ingestion for RAPTA-C . ( B ) Body weight (g) measured at treatment day 12. ( C ) Microvessel density (MVD) analysis in combination III treated tumors represented as the number of vessels per mm 2 of vascularized tumor area and represented as percentage of control. ( D ) Analysis of staining for the proliferation marker Ki67 in combination III treated tumors shown as the density of Ki67 positive nuclei per image field and represented as percentage CTRL. ( E ) Representative images of IHC staining for the endothelial cell marker CD31 (brown) counter-stained with haematoxylin (purple/blue). ( F ) Representative images of IHC staining for Ki67 (brown/orange) counter-stained with haematoxylin (purple/blue). III indicates the combination erlotinib 10 mg/kg/day + RAPTA-C 100 mg/kg/day in all graphs. Error bars represent SEM and *P < 0.05 indicates significance vs. CTRL. Statistical analysis was performed using a one-way ANOVA with post-hoc Tukey’s multiple comparison test ( A ), or an unpaired, one-tailed t-test with Welch’s correction.

Article Snippet: Sections were stained separately for proliferation marker with primary antibodies against Ki67 (rabbit anti-human Ki67, 1:100, clone SP6; Thermo Scientific) and subsequent incubation with secondary swine anti-rabbit biotinylated antibodies (1:200; Dako, Glostrup, Denmark).

Techniques: Control, Clinical Proteomics, Staining, Marker, Immunohistochemistry, Comparison, One-tailed Test

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Rosmarinic inhibits cell proliferation, invasion and migration via up-regulating miR-506 and suppressing MMP2/16 expression in pancreatic cancer.

doi: 10.1016/j.biopha.2019.108878

Figure Lengend Snippet: Fig. 7. Rosmarinic acid inhibited in vivo tumor growth in the xenograft mice model. (A) The tumor volume in the xenograft mice received different doses of rosmarinic acid treatment was determined every 5 days for 30 days. (B) The weight of the dissected tumors from the xenograft mice with different doses of rosmarinic acid treatments was determined. (C–E) The miR-506 expression and MMP2/16 mRNA expression in the dissected tumor tissues were determined by qRT-PCR assay. (F) The MMP2/16 protein expression in the dissected tumor tissues were determined by western blot assay. (G) The Ki-67 expression in the dissected tumor tissues were determined by immunohistochemistry. Scale bar = 100 μm. RA = rosmarinic acid; n = 5; *P < 0.05, **P < 0.01 and ***P < 0.001.

Article Snippet: For the determination of Ki-67 expression, the paraffin-embedded tumor tissues were immunostaining with primary rabbit anti-human Ki67 antibodies (Cell Signaling Technology, Danvers, USA) according to previous studies [24].

Techniques: In Vivo, Expressing, Quantitative RT-PCR, Western Blot, Immunohistochemistry

Journal: STAR Protocols

Article Title: Simplified mass cytometry protocol for in-plate staining, barcoding, and cryopreservation of human PBMC samples in clinical trials

doi: 10.1016/j.xpro.2022.101362

Figure Lengend Snippet:

Article Snippet: Anti-Ki-67 (B56)-168Er—100 Tests (1:100) , Fluidigm , Cat#3168007B.

Techniques: Recombinant, Saline, Staining, Blocking Assay, Isolation, Software, Cell Counting, Cytometry