Journal: Oncotarget

Article Title: Cabazitaxel operates anti-metastatic and cytotoxic via apoptosis induction and stalls brain tumor angiogenesis

doi: 10.18632/oncotarget.9439

Figure Lengend Snippet: A. Isolated primary hippocampal neurons and astrocytes were treated with 1, 5 and 10 nM cabazitaxel for 3 days. Neurons and astrocyte were stained with anti Tuj-1 (green) and GFAP (red) respectively. Scale bar represents 100 μm. B. Quantification of Tuj-1 (β-III tubulin) and GFAP immunostaining ( n = 3 ). Statistical analysis was performed with One-way ANOVA (mean is given ± s.e.m.).

Article Snippet: Primary β-tubulin III (Tuj-1) (Promega) and GFAP antibodies (Dako) were diluted in 0.4% Triton-X (Sigma-Aldrich) + 3% FCS as the blocking solution and subsequently incubated with the samples for overnight at 4°C.

Techniques: Isolation, Staining, Immunostaining

Journal: Frontiers in Pharmacology

Article Title: Leishmanicidal and immunomodulatory properties of Brazilian green propolis extract (EPP-AF ® ) and a gel formulation in a pre-clinical model

doi: 10.3389/fphar.2023.1013376

Figure Lengend Snippet: EPP-AF ® treatment induces HO-1 expression. BMDM were infected with L. amazonensis and treated with 25 μg/mL of EPP-AF ® as described in Materials and Methods . After 48 h, cell extract was collected to evaluate protein levels of (A) HO-1 by western blotting, with anti-β-Actin antibody used as an endogenous control. In a parallel assay, after 48 h of treatment, the culture medium was changed and (B) viable promastigotes were counted after 6 days in the presence or absence of the heme oxygenase inhibitor tin protoporphyrin (SnPP) and/or propolis. In (A) , Western blotting results are presented as a single experiment performed in quintuplicate, representative of three independent experiments. In (B) , bars represent mean parasite burden (± SD) from experiments performed in quintuplicate, representative of two independent experiments. The Kruskal-Wallis test, followed by Dunn’s post-test, were used for multiple comparisons, while the Mann-Whitney test was employed to compare two groups (* p < 0.05). La: Leishmania amazonensis ; UN: Uninfected; EPP-AF ® : Brazilian green propolis extract at 25 μg/mL.

Article Snippet: Primary β-actin, pERK and ERK antibodies were obtained from Cell Signaling (Danvers, MA, United States), while Cox-2 and iNOS antibodies were purchased from Calbiochem (San Diego, CA, United States).

Techniques: Expressing, Infection, Western Blot, Control, MANN-WHITNEY

Journal: Frontiers in Pharmacology

Article Title: Leishmanicidal and immunomodulatory properties of Brazilian green propolis extract (EPP-AF ® ) and a gel formulation in a pre-clinical model

doi: 10.3389/fphar.2023.1013376

Figure Lengend Snippet: EPP-AF ® modulates iNOS and COX-2 enzyme expression, without impacting in arginase or ODC. BMDM were infected or not with L. amazonensis, stimulated or not with IFN-γ, and treated with EPP-AF ® at 25 μg/mL, as described in Materials and Methods . After 48 h of treatment, the cell extract was collected to evaluate protein expression by Western Blotting. (A) Arginase and ornithine decarboxylase (ODC), (B) iNOS and COX-2 (anti-β-Actin antibody used as an endogenous control). The levels of (C) nitrite and (D) PGE 2 were evaluated in cell culture supernatants. Bars represent ±SD of experiments performed in quintuplicate, representative of three or one independent experiment(s). Western blotting results are presented as an experiment performed in quintuplicate, representative of two (Arginase-1 and ODC) or three (iNOS and COX-2) independent experiments. The Kruskal-Wallis test, followed by Dunn’s post-test, were used for multiple comparisons (* p < 0.05; ** p < 0.01; # p < 0.05 in the comparison of uninfected condition (light gray bar) versus infected (black bar); + p < 0.05 in the comparison of the NS398 exposure condition versus levels of PGE2 in infected cells. La: Leishmania amazonensis ; NS: Not Stimulated; UN: Uninfected and IFN-y stimulated; EPP-AF ® : Brazilian green propolis extract at 25 μg/mL.

Article Snippet: Primary β-actin, pERK and ERK antibodies were obtained from Cell Signaling (Danvers, MA, United States), while Cox-2 and iNOS antibodies were purchased from Calbiochem (San Diego, CA, United States).

Techniques: Expressing, Infection, Western Blot, Control, Cell Culture, Comparison

Journal: Frontiers in Pharmacology

Article Title: Leishmanicidal and immunomodulatory properties of Brazilian green propolis extract (EPP-AF ® ) and a gel formulation in a pre-clinical model

doi: 10.3389/fphar.2023.1013376

Figure Lengend Snippet: EPP-AF ® treatment activates ERK 1/2. BMDM were infected or not with L. amazonensis and treated with EPP-AF propolis extract at 25 μg/mL for 15, 30 or 45 min. After each treatment time, cell extracts were collected and analyzed by Western Blotting to evaluate the protein levels of (A) phosphorylated (pERK) and total (Total ERK) ERK. Anti-β-Actin antibody was used as an endogenous control. Levels of (B) TNF-α produced in the presence or absence of the ERK 1/2 inhibitor (PD98059) and/or propolis and (C) infection rate obtained under the same conditions. Bars represent ± SD of a representative experiment out of two independent experiments, performed in quintuplicate (Western blotting assay) or one representative experiment performed in quintuplicate (TNF-α and parasite burden assays). The Kruskal-Wallis test, followed by Dunn’s post-test were used for multiple comparisons, and Mann-Whitney testing was applied for comparisons between two groups (* p < 0.05 and ** p < 0.01); For comparisons of TNF-α (C) levels between uninfected (dark gray bar) and infected (black bar) conditions, ## p < 0.01. La: Leishmania amazonensis , UN: Uninfected, PD: PD98059 and EPP-AF ® : Brazilian green propolis extract at 25 μg/mL.

Article Snippet: Primary β-actin, pERK and ERK antibodies were obtained from Cell Signaling (Danvers, MA, United States), while Cox-2 and iNOS antibodies were purchased from Calbiochem (San Diego, CA, United States).

Techniques: Infection, Western Blot, Control, Produced, MANN-WHITNEY

Journal: Scientific reports

Article Title: Probiotics may delay the progression of nonalcoholic fatty liver disease by restoring the gut microbiota structure and improving intestinal endotoxemia.

doi: 10.1038/srep45176

Figure Lengend Snippet: Figure 6. Effects of probiotics on intestinal barrier integrity in NAFLD models. (A) Representative images of tight junctions in the jejunal mucosa (transmission electron microscopy, x5000) (B) The expression of occludin protein in the intestinal mucosa was detected with β-actin as a loading control by western blotting. Data represent the mean ± SD of each group. *P < 0.05 compared to the control group; **P < 0.01 compared to the control group; ***P < 0.001 compared to the control group; #P < 0.05 compared to the model group; ##P < 0.01 compared to the model group; ###P < 0.001 compared to the model group.

Article Snippet: Primary occludin (ab31721) antibody was from Cell Signaling Technology (Cambridge, MA, USA), and primary β -actin (66009-1) antibody was from Proteintech (Chicago, USA).

Techniques: Probiotics, Transmission Assay, Electron Microscopy, Expressing, Control, Western Blot