Journal: Toxicology and applied pharmacology

Article Title: Role of the PI3K/Akt pathway in cadmium induced malignant transformation of normal prostate epithelial cells.

doi: 10.1016/j.taap.2020.115308

Figure Lengend Snippet: Cell phenotype and tumor formation in nude mice. Parental (RWPE1/PWR1E) and cadmium-exposed (Cd-RWPE1/Cd-PWR1E) cells under the reflected light microscope (A). Sketch showing injection scheme (B). Tumor formation by Cd-RWPE1 cells (C-D). Batch matched parental RWPE1 cells did not result in any tumors (D). Tumor volume over the course of experiment (D). There were four replicates for each cell line. Error bars ±SE.

Article Snippet: Human prostate epithelial cell lines and their culture Human normal prostate epithelial cell lines RWPE1 and PWR1E were purchased from American Type Culture Collection (ATCC) (Manassas, VA).

Techniques: Light Microscopy, Injection

Journal: Toxicology and applied pharmacology

Article Title: Role of the PI3K/Akt pathway in cadmium induced malignant transformation of normal prostate epithelial cells.

doi: 10.1016/j.taap.2020.115308

Figure Lengend Snippet: Wound healing assay. Percent artificial wound closure after 24 hours (A; G). Representative pictures of the artificial wound at 0 and 24 hours of cell culture (B; H). Error bar ±SD. Chemotactic cell migration and invasion assay. Average percent migration (C; I) and invasion (E; K) in Cd-exposed (Cd-RWPE1 and Cd-PWR1E) cells compared to their normal parental (RWPE1 and PWR1E) controls. Data are represented as the mean ±SD from at least three replicates. Representative pictures of migration (D; J) and invasion (F; L) in both cell lines.

Article Snippet: Human prostate epithelial cell lines and their culture Human normal prostate epithelial cell lines RWPE1 and PWR1E were purchased from American Type Culture Collection (ATCC) (Manassas, VA).

Techniques: Wound Healing Assay, Cell Culture, Migration, Invasion Assay

Journal: Toxicology and applied pharmacology

Article Title: Role of the PI3K/Akt pathway in cadmium induced malignant transformation of normal prostate epithelial cells.

doi: 10.1016/j.taap.2020.115308

Figure Lengend Snippet: Differential expression by RT2 profiler array, real-time PCR and immunoblot analyses of the PI3K/Akt signaling pathway. Scatter plot representing the array results. Upregulated genes are indicated by red points and downregulated genes are in green (A). Heat Map shows the fold changes in expression of 84 PI3K/Akt pathway genes in Cd-RWPE1 vs. RWPE1 cells (B). The complete gene names, location on array and fold changes are indicated in Supplemental Table 1. TaqMan gene expression (C-I) and immunoblot (J) analyses of selected PI3K/Akt pathway genes. Graphical representation of the relative fold upregulation in protein levels based on the average intensity of the target proteins in all replicates and normalized to their endogenous controls (K). Error bars ±SD. Experiments were performed three times and each time there were three replicates in TaqMan assays. For RT2 profiler array RNA from three biological replicates was pooled together for the cDNA synthesis for all four cell lines.

Article Snippet: Human prostate epithelial cell lines and their culture Human normal prostate epithelial cell lines RWPE1 and PWR1E were purchased from American Type Culture Collection (ATCC) (Manassas, VA).

Techniques: Quantitative Proteomics, Real-time Polymerase Chain Reaction, Western Blot, Expressing, Gene Expression, cDNA Synthesis

Journal: Toxicology and applied pharmacology

Article Title: Role of the PI3K/Akt pathway in cadmium induced malignant transformation of normal prostate epithelial cells.

doi: 10.1016/j.taap.2020.115308

Figure Lengend Snippet: Significantly up- (> 2 fold) and down-regulated ( <0.5) genes in Cd exposed (Cd-RWPE1; Cd-PWR1E) compared to parental normal (RWPE1; PWR1E) cells.

Article Snippet: Human prostate epithelial cell lines and their culture Human normal prostate epithelial cell lines RWPE1 and PWR1E were purchased from American Type Culture Collection (ATCC) (Manassas, VA).

Techniques:

Journal: Toxicology and applied pharmacology

Article Title: Role of the PI3K/Akt pathway in cadmium induced malignant transformation of normal prostate epithelial cells.

doi: 10.1016/j.taap.2020.115308

Figure Lengend Snippet: Cd-RWPE1 is denoted as “1” and parental normal RWPE1 as “0” in the plots. Five overlapping pathways are enriched in Cd-RWPE1 vs. Parental RWPE1 as control. These pathways are KEGG_Prostate cancer Pathway (A). KEGG_Apoptosis (B). KEGG_MAPK (C). KEGG_Pathways in cancer (D). KEGG_ERBB (E) pathways. Same pathways were enriched in Cd-PWR1E vs. normal parental PWR1E cells (Supplemental Figures 1 and 2).

Article Snippet: Human prostate epithelial cell lines and their culture Human normal prostate epithelial cell lines RWPE1 and PWR1E were purchased from American Type Culture Collection (ATCC) (Manassas, VA).

Techniques: Control

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Ginkgolic acid improves bleomycin-induced pulmonary fibrosis by inhibiting SMAD4 SUMOylation

doi: 10.1155/2022/8002566

Figure Lengend Snippet: Ginkgolic acid (GA) mediated TGF- β 1-induced SMAD4 SUMOylation in a mouse model of pulmonary fibrosis. (a) Lung tissues of wild-type (WT) mice in the different administration/treatment groups were used in Co-IP experiments with anti-SUMO1and anti-SMAD4 antibodies, followed by western blot analysis with the indicated antibodies (input). The results showed that TGF could reverse the inhibitory effect of GA on the interaction between SUMO1 and SMAD4, not SMAD2/3. (b) The effect of GA on SMAD4 SUMOylation in WT mice was analyzed by immunofluorescence. (c) Lung tissues of SENP1-KO transgenic mice in the different administration/treatment groups were used in Co-IP experiments as in (a). (d) The effect of SENP1 on the SUMOylation of SMAD4 was investigated by Co-IP in A549 cells transfected with the indicated plasmids.

Article Snippet: Adenocarcinomic human alveolar basal epithelial cells A549 cells were obtained from ATCC (Manassas, VA, USA).

Techniques: Co-Immunoprecipitation Assay, Western Blot, Immunofluorescence, Transgenic Assay, Transfection

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Ginkgolic acid improves bleomycin-induced pulmonary fibrosis by inhibiting SMAD4 SUMOylation

doi: 10.1155/2022/8002566

Figure Lengend Snippet: SENP1 mediated the deSUMOylation of SMAD4 in response to TGF- β 1-induced EMT. A549 cells were either transfected with His-SENP1 or treated with BLM, GA, NAC (TGF- β 1 scavenger agonist), or them in combination. (a) The morphological changes of cells in each group were observed under light microscopy. (b) Western blot (WB) analysis of the levels of key EMT-related proteins. (c) Vimentin expression was detected by immunofluorescence to determine the degree of pulmonary fibrosis. Data are presented as the mean ± SD. n = 6, ∗ P < 0.05.

Article Snippet: Adenocarcinomic human alveolar basal epithelial cells A549 cells were obtained from ATCC (Manassas, VA, USA).

Techniques: Transfection, Light Microscopy, Western Blot, Expressing, Immunofluorescence

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Ginkgolic acid improves bleomycin-induced pulmonary fibrosis by inhibiting SMAD4 SUMOylation

doi: 10.1155/2022/8002566

Figure Lengend Snippet: SENP1 mediated the deSUMOylation of SMAD4 in response to TGF- β 1-induced ROS generation. A549 cells were either transfected with His-SENP1 or treated with BLM, GA, TGF- β 1, or them in combination. (a) The fluorescence intensity of 2′,7′-dichlorofluorescein (DCF) in cells of each group was detected using a reactive oxygen species (ROS) assay kit to show ROS levels in cells of each treatment group. (b) WB analysis of the key oxidases that triggered the ROS outbreak to clarify the role of SMAD4 SUMOylation and SENP1 in TGF- β 1-induced ROS generation. (c) A549 cells transfected with siRNA-SENP1 or treated with BLM, GA, TGF- β 1, NAC, or them in combination were subjected to WB analysis to detect the levels of key protein involved in EMT. Data are presented as the mean ± SD. n = 6, ∗ P < 0.05.

Article Snippet: Adenocarcinomic human alveolar basal epithelial cells A549 cells were obtained from ATCC (Manassas, VA, USA).

Techniques: Transfection, Fluorescence, ROS Assay