Journal: Pharmaceuticals (Basel, Switzerland)

Article Title: Skimmianine Modulates Tumor Proliferation and Immune Dynamics in Breast Cancer by Targeting PCNA and TNF-α.

doi: 10.3390/ph18050756

Figure Lengend Snippet: Figure 2. Cross-sections of mammary glands in (A) control, (B) DMBA, and (C) DMBA + skimmianine groups. (D) Semi-quantitative measurement of PCNA expression. Ductal structures (d), connective tissue (ct), malignant cells (*), fatty tissue (f), arrow: negative expression, arrowhead: positive expression. Magnification: 20× (low), 40× (insets).

Article Snippet: An immunohistochemistry with PCNA (sc-25280 Santa Cruz, Dallas, TX, USA) ve TNFα (sc-52746 Santa Cruz USA) was performed according to the biotin–streptavidin peroxidase complex method described previously [60].

Techniques: Control, Expressing

Journal: Pharmaceuticals (Basel, Switzerland)

Article Title: Skimmianine Modulates Tumor Proliferation and Immune Dynamics in Breast Cancer by Targeting PCNA and TNF-α.

doi: 10.3390/ph18050756

Figure Lengend Snippet: Figure 4. Protein–protein interaction (PPI) network and functional enrichment analysis of skimmianine-associated PCNA (A) and TNF-α (B) targets. PPI network visualization illustrating hub proteins ranked by importance using the MCC algorithm, where red represents the highest-ranked hub proteins and yellow indicates lower-ranked hub proteins. Functional annotation of shared targets presented as a bar chart, with Reactome pathways ranked by ascending adjusted p-values (p < 0.05). NLR: Leucine Rich Repeat Containing Receptor.

Article Snippet: An immunohistochemistry with PCNA (sc-25280 Santa Cruz, Dallas, TX, USA) ve TNFα (sc-52746 Santa Cruz USA) was performed according to the biotin–streptavidin peroxidase complex method described previously [60].

Techniques: Functional Assay

Journal: Pharmaceuticals (Basel, Switzerland)

Article Title: Skimmianine Modulates Tumor Proliferation and Immune Dynamics in Breast Cancer by Targeting PCNA and TNF-α.

doi: 10.3390/ph18050756

Figure Lengend Snippet: Figure 5. Immunological analysis of PCNA and TNF-α in breast cancer. (A) Correlation of PCNA and TNF-α expression with molecular and immune subtypes, showing their increased expression in the basal subtype and their highest expression in the C2 (IFN-γ dominant) immune subtype (CPM: counts per million). ImmuneScore analysis assessing the association between immune modulation and key hub proteins derived from the skimmianine-associated PCNA (B) and TNF-α (C) PPI networks, identifying significantly correlated proteins (p < 0.05).

Article Snippet: An immunohistochemistry with PCNA (sc-25280 Santa Cruz, Dallas, TX, USA) ve TNFα (sc-52746 Santa Cruz USA) was performed according to the biotin–streptavidin peroxidase complex method described previously [60].

Techniques: Expressing, Derivative Assay

Journal: Pharmaceuticals (Basel, Switzerland)

Article Title: Skimmianine Modulates Tumor Proliferation and Immune Dynamics in Breast Cancer by Targeting PCNA and TNF-α.

doi: 10.3390/ph18050756

Figure Lengend Snippet: Figure 6. Correlation of skimmianine-associated PCNA and TNF-α PPI hub proteins with tumor immune infiltration. Heatmap analysis depicting the correlation between skimmianine-associated PCNA (ATM, CDK2, CDK4, HDAC1, MDM2) and TNF-α (BCL2, CASP3, IL2, MMP9, TNF-α) hub proteins with six major immune cell infiltrates in breast cancer. In the heatmaps, green regions denote correlation coefficients, whereas black regions represent p-values, with significance levels indicated as * p < 0.05, ** p < 0.01, *** p < 0.005, and **** p < 0.001.

Article Snippet: An immunohistochemistry with PCNA (sc-25280 Santa Cruz, Dallas, TX, USA) ve TNFα (sc-52746 Santa Cruz USA) was performed according to the biotin–streptavidin peroxidase complex method described previously [60].

Techniques:

Journal: Cell death & disease

Article Title: FGF23 ameliorates ischemia-reperfusion induced acute kidney injury via modulation of endothelial progenitor cells: targeting SDF-1/CXCR4 signaling.

doi: 10.1038/s41419-021-03693-w

Figure Lengend Snippet: Fig. 1 FGF23 ameliorates kidney ischemia-reperfusion (IR) injury in mice. A, F Plasma levels of iFGF23 were determined using ELISA kit after 30 min of FGF23 protein (A) or 24 h of pFGF23 plasmid (F) injection. Bars on graph are ±SEM (n = 3). ***p < 0.001 versus control mice by t test. B, C Recombinant mouse FGF23 (rmFGF23) were injected into C57BL/6 mice via tail-vein 30 min before IR injury. G, H FGF23 expression plasmid (pFGF23) was injected into C57BL/6 mice via tail-vein 24 h before IR injury. Serum blood urea nitrogen (BUN) (B, G) and creatinine (CRE) (C, H) were measured after 48 h of reperfusion. Bars on graphs are ± SEM. *p < 0.05 by t test, n = 9. D, I Kidneys were harvested after 48 h of reperfusion for H&E staining from FGF23 protein-injected (D) or pFGF23-injected mice (I). Necrotic tubules were marked with asterisks. Scale bars, 50 μm (D), 100 μm (I). Kidney injury score was graded using a semi- quantitative scale in ten random fields for each sample (n = 9). Bars on graph are ±SEM. ***p < 0.001 by t test. E, J Kidneys were harvested after 48 h of reperfusion immunohistochemistry with CD31 antibody from FGF23 protein-injected (E) or pFGF23-injected mice (J). Scale bars, 20 μm (E), 100 μm (J). The percentage of CD31 positive cells was counted in ten random fields for each sample (n = 9). Bars on graph are ±SEM. **p < 0.01, ***p < 0.001 by t test.

Article Snippet: The following commercial primary antibodies were used for immunohistochemistry: PCNA (Cell Signaling, #13110, 1:2000), p-Akt (Cell Signaling, #4060, 1:50), or CD31 (Cell Signaling, #77699, 1:100).

Techniques: Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Plasmid Preparation, Injection, Control, Recombinant, Expressing, Staining, Immunohistochemistry

Journal: American Journal of Translational Research

Article Title: Tanshinone IIA regulates fibroblast proliferation and migration and post-surgery arthrofibrosis through the autophagy-mediated PI3K and AMPK-mTOR signaling pathway

doi:

Figure Lengend Snippet: Tan IIA alleviated post-surgery arthrofibrosis in vivo. (A) Immunofluorescence staining of vimentin and FSP-1 in vivo showed a linear signal, with vimentin (red) and FSP-1 (green) localized in a fibrous structure. DAPI (blue) was used to localize the nuclei; scale bar, 100 μm. (B) HE staining was used to detect scar density, and Masson and Sirius Red (SR) staining was used to indicate the variation in collagen synthesis in vivo; scale bar, 50 μm. (C) Statistical analysis of collagen synthesis with SR staining. (D) IHC staining of PCNA, tubulin and LC3B in vivo, and (E, F) the results of statistical analyses of the positive cell rates; scale bar, 60 μm; (G) Chemical structure of Tan IIA. Data are shown as the means ± standard deviations (SDs) from three independent samples; *P < 0.05 vs. control.

Article Snippet: Immunohistochemistry (IHC) analysis PCNA (bs-2007R, Bioss), tubulin (ba-20694R, Bioss) and LC3B (NB100-2220, Novus, Minnesota, USA) protein expression in intra-articular fibrosis scar tissue was detected with IHC staining.

Techniques: In Vivo, Immunofluorescence, Staining, Immunohistochemistry

Journal: BMC Genomics

Article Title: EPIG-Seq: extracting patterns and identifying co-expressed genes from RNA-Seq data

doi: 10.1186/s12864-016-2584-7

Figure Lengend Snippet: PCNA expression. a Gene expression of PCNA in TCGA normal and breast cancer samples. The x-axis denotes the breast cancer tumor subtype. The y-axis is the average of the log base 2 ratio of PCNA in each tumor subtype relative to the average of the normal samples. Standard error bars are shown for each data point. b PCNA protein immunohistochemistry staining of normal breast tissue with benign adenomas from a female age 23 (ID: 2773) and using the HPA030522 antibody. c PCNA protein immunohistochemistry staining of breast cancer tissue (ductal carcinoma) from a female age 55 (ID: 2773) and using the HPA030522 antibody

Article Snippet: Interestingly, the Human Protein Atlas [ ] contains immunohistochemistry staining of the PCNA protein (antibody HPA030522) in normal breast tissue (Fig. ) versus the overexpression in breast cancer ductal carcinoma (Fig. ) implicating the abundance of the protein in breast cancer tissue as a potential biomarker [ ].

Techniques: Expressing, Gene Expression, Immunohistochemistry, Staining