Journal: The Journal of Biological Chemistry

Article Title: Peroxiredoxin IV plays a critical role in cancer cell growth and radioresistance through the activation of the Akt/GSK3 signaling pathways

doi: 10.1016/j.jbc.2022.102123

Figure Lengend Snippet: Examination of Prx4 protein levels in patient specimens of prostate cancer by tissue microarray or established prostate cancer cell lines by Western blotting. A , tissue microarray slides and representative images of anti-Prx4 staining of prostate normal and adenocarcinoma. Positive anti-Prx4 staining shows in dark brown and counter staining for nuclear shows light blue . The scale bars represent 5 mm (whole slide), 500 μm (individual tumor), and 100 μm (zoomed in). B and C , anti-Prx4 staining intensity were quantitatively scored by board-certified pathologist and data were compared between prostate normal (n = 50) and adenocarcinoma (n = 66) ( B ), as well as between prostate normal and cancer with different levels of Gleason grading ( C ). D , examination of Prx4 levels in prostate derived nontumorigenic or tumor cell lines by Western blotting. The bar graph on the right shows quantitative data from three independent blots. ∗ p < 0.05 by two-way ANOVA. Prx, peroxiredoxin.

Article Snippet: Human prostate normal and prostate cancer tissue microarray slides were commercially obtained, including BNS19011 and PR803d (US Biomax).

Techniques: Microarray, Western Blot, Staining, Derivative Assay

Journal: International Journal of Molecular Sciences

Article Title: Macrophages Cytokine Spp1 Increases Growth of Prostate Intraepithelial Neoplasia to Promote Prostate Tumor Progression

doi: 10.3390/ijms23084247

Figure Lengend Snippet: Increased infiltrating macrophages in the prostate intraepithelial neoplasia were M2 subtype. ( A ) Human tissue samples of prostate cancer containing the area of prostate intraepithelial neoplasia (PIN) were immunostained with the antibody of CD68, a pan marker for macrophages (top panel, and middle panel: zoom-in areas). Hematoxylin and eosin (H & E) staining was carried out to visualize the morphology of the prostate cancer tissues (bottom panel). Scale bar: 25 µm. ( B , C ) Raw 264.7 macrophages cultured with either control media or Pr111 PIN complete media were fixed, permeabilized, and immunostained with antibodies for markers of pan macrophages, F4/80; M1 macrophages including iNOS ( B ) and CD38 ( C ); M2 macrophages including Ym1 ( B ) and CD206 ( C ). Scale bar: 20 µm. ( D ) Human tissue samples of PIN were immunostained with antibodies of CD68 and CD206 for the assessment of all macrophages (green) and M2 macrophages (red), respectively. Nuclei were labeled by DAPI and visualized. Scale bar: 50 µm. The image shown here is representative of at least 3 independent experiments or tissue samples.

Article Snippet: Human prostate tissue slides, including normal or cancer tissue samples, were obtained from BioChain (Newark, CA, USA), US Biomax (Derwood, MD, USA), and Cooperative Human Tissue Network (CHTN).

Techniques: Marker, Staining, Cell Culture, Control, Labeling

Journal: International Journal of Molecular Sciences

Article Title: Macrophages Cytokine Spp1 Increases Growth of Prostate Intraepithelial Neoplasia to Promote Prostate Tumor Progression

doi: 10.3390/ijms23084247

Figure Lengend Snippet: Expression of Spp1 receptors in PIN cells. ( A ) Cell lysates from murine PIN Pr111 cells grown on Matrigel in 3D treated with recombinant Spp1 or control for 72 h were subjected to immunoblotting for assessment of the Spp1 receptors, including integrin αvβ3, integrin β1, and CD44. Long exposure: longer time to catch the emitted signal from the WB membrane on the X-ray film. ( B – E ) Human tissue samples of normal prostate and prostate cancer containing PIN areas were immunostained for the expression of CD44 ( B ), integrin αv ( C ), integrin β1 ( D ), and integrin β3 ( E ). Scale bar: 50 µm. the image shown here is representative of at least 3 independent experiments or tissue samples.

Article Snippet: Human prostate tissue slides, including normal or cancer tissue samples, were obtained from BioChain (Newark, CA, USA), US Biomax (Derwood, MD, USA), and Cooperative Human Tissue Network (CHTN).

Techniques: Expressing, Recombinant, Control, Western Blot, Membrane

Journal: International Journal of Molecular Sciences

Article Title: Macrophages Cytokine Spp1 Increases Growth of Prostate Intraepithelial Neoplasia to Promote Prostate Tumor Progression

doi: 10.3390/ijms23084247

Figure Lengend Snippet: Macrophage Spp1 activated Akt and JNK signaling in PIN cells. ( A – C ) Pr111 cells cultured on Matrigel in 3D were stimulated with either control/ddH 2 O or recombinant Spp1 for 72 h. Cell lysates were collected and subjected to immunoblotting to examine the proteins of interest, including p-Akt, Akt, p-ERK, and ERK ( A ); p-JNK, JNK, p38 MAPK, p38 MAPK, p-Src, and Src ( B ); and IκBα ( C ). The relative intensity fold change was calculated by the ratio of phosphorylated protein levels over the corresponding total protein levels and set as 1 for the control in each case. Each image shown is representative of 3–5 independent experiments. ( D ) Human tissue samples of normal prostate and prostate cancer containing PIN areas were immunostained for phospho-Akt expression. Scale bar: 25 µm. n = 3.

Article Snippet: Human prostate tissue slides, including normal or cancer tissue samples, were obtained from BioChain (Newark, CA, USA), US Biomax (Derwood, MD, USA), and Cooperative Human Tissue Network (CHTN).

Techniques: Cell Culture, Control, Recombinant, Western Blot, Expressing