Journal: Biology of reproduction

Article Title: Members of the Toll-like receptor family of innate immunity pattern-recognition receptors are abundant in the male rat reproductive tract.

doi: 10.1095/biolreprod.106.059410

Figure Lengend Snippet: FIG. 2. Immunoblot analysis of TLR proteins in male reproductive organs. Aliquots (100 lg) of cytoplasmic protein extracts were separated by PAGE, electroblotted, and blots probed with anti-TLR antibodies followed by enhanced chemiluminescence detection using Pierce Super- Signal West Pico or Pierce SuperSignal West Femto (*F) substrate. Representative results are shown (n ¼ 3–5 rats). þ, control rat tissue extracts from spleen, TLRs 1–7; lung, TLR8; and small intestine, TLR9. þ Ext, positive-control whole cell lysates used were Raw 264 Abelson transformed macrophages (TLRs 1, 2–6, and 8–10); SW480 colorectal adenocarcinoma (TLR2); Daudi cell extract (TLR7); Ramos cell lysate (TLR10) and mouse heart whole cell lysate (TLR11). Blots were stripped and reprobed with anti-actin monoclonal antibody to detect actin as a loading control. Representative results are shown (n ¼ 3–5 rats) for blots exposed to film for the same length of time when using equivalent chemiluminescent substrate.

Article Snippet: Mouse RAW 264.7 þ LPS/IFN-c Abelson-transformed macrophage whole cell lysate (sc-24767), SW480 human colorectal adenocarcinoma whole cell lysate (sc-2219), and Daudi cell (Burkitt lymphoma) whole cell lysate (sc2415) were purchased from Santa Cruz Biotechnology.

Techniques: Western Blot, Control, Positive Control, Transformation Assay

Journal: Acta Pharmaceutica Sinica. B

Article Title: CRISPR-Cas9 gene editing strengthens cuproptosis/chemodynamic/ferroptosis synergistic cancer therapy

doi: 10.1016/j.apsb.2024.05.029

Figure Lengend Snippet: In vitro distribution and tumor-combatting effect of RNP@Cu 2 O@SPF. (A) The viability of cancer and normal cells treated with RNP@Cu 2 O@SPF for 24 h in the dark ( n = 6). (B) CLSM images of SW480 cells incubated with RNP@Cu 2 O@SPF or RNP@Cu 2 O@S (FITC-labelled RNP, RITC-labelled silica). Scale bar, 20 μm. (C) FCM analysis showcasing transfection rates in SW480 cells after 4 h of immersion with PBS, RNP@Cu 2 O@S, or RNP@Cu 2 O@SPF. WB assay (D) and fluorescence imaging (E) of ATP7A expression in SW480 cells treated with RNP@Cu 2 O@S or RNP@Cu 2 O@SPF. Scale bar, 100 μm. (F) Relative copper contents in SW480 cells after a 24 h treatment with Cu 2 O@SPF or RNP@Cu 2 O@SPF ( n = 6). (G) Fluorescence microscopy images of ROS generation in SW480 cells treated with PBS, Cu 2 O@SPF(40 μg/mL), RNP@Cu 2 O@SPF (40 μg/mL), and RNP@Cu 2 O@SPF (80 μg/mL) for 4 h and stained with DCFH-DA to indicate the ROS generation. Scale bar, 50 μm. (H) GSH/GSSG ratio in SW480 cells treated with different concentrations of RNP@Cu 2 O@SPF (25, 50, 100 μg/mL) for 6 h ( n = 6). The data are shown as mean ± SD. ns, no significant; ∗ P < 0.05, ∗∗ P < 0.01.

Article Snippet: The Cell Copper (Cu) Colorimetric Assay Kit (Elabscience Biotechnology, Wuhan, China) was selected to detect the copper ion content in SW480 cells, and the BCA Protein Colorimetric Assay Kit (Elabscience Biotechnology, Wuhan, China) was also used to determine the total protein concentration.

Techniques: In Vitro, Incubation, Transfection, Fluorescence, Imaging, Expressing, Microscopy, Staining

Journal: Acta Pharmaceutica Sinica. B

Article Title: CRISPR-Cas9 gene editing strengthens cuproptosis/chemodynamic/ferroptosis synergistic cancer therapy

doi: 10.1016/j.apsb.2024.05.029

Figure Lengend Snippet: Anti-tumor mechanisms studies of RNP@Cu 2 O@SPF. (A) The schematic diagram of cell death mechanisms of RNP@Cu 2 O@SPF nanomedicine-mediated synergetic therapy. (B) FCM analysis with Annexin V-FITC/PI dual labels on SW480 cells after 24 h of incubation with different nanoparticles. (C) The percentages of cells undergoing apoptosis (%) in different groups ( n = 3). (D) Viability of SW480 cells grown in media containing either glucose or galactose treated with RNP@Cu 2 O@SPF (ratio 1:1) ( n = 6). (E) Viability of SW480 cells pretreated with 0.1 mmol/L rotenone (Rot), 0.1 mmol/L antimycin A (anti-A), or 1 mmol/L FCCP and then treated with RNP@Cu 2 O@SPF ( n = 6). (F) Viability of SW480 cells post-treated with Antimycin A or UK5099 under different conditions (incubated with various nanoparticles) ( n = 6). (G) Viability of SW480 cells treated with tetrathiomolybdate in different concentrations of RNP@Cu 2 O@SPF (50, 100 μg/mL) ( n = 6). (H) Western blot analysis of DLAT-oligomers, DLAT, and Tubulin expression levels in SW480 cells after treatment with different nanoparticles. (I) Western blot analysis of lipoylated proteins, FDX1, LIAS, ACO-2, SDHB9, and Tubulin expression levels in SW480 cells after treatment with different nanoparticles. (J) Fluorescence depiction of GPX4 levels in SW480 cells post RNP@Cu 2 O@SPF treatment at different dosages (25, 50, and 100 μg/mL) for 24 h. Scale bar, 25 μm. (K) LPO contents in SW480 cells treated with RNP@Cu 2 O@SPF at different concentrations (25, 50, and 100 μg/mL) for 24 h ( n = 6). (L) Survival rates of SW480 cells after 24 h of incubation with solutions of PBS, RNP@Cu 2 O@SPF, PBS+DFO, or RNP@Cu 2 O@SPF+DFO ( n = 6). (M) WB assay of GPX4, ACSL4, and Tubulin expression in SW480 cells treated with RNP@Cu 2 O@SPF at varied concentrations (25, 50, and 100 μg/mL) for 24 h. The data are shown as mean ± SD. ns, not significant; ∗ P < 0.05, ∗∗ P < 0.01.

Article Snippet: The Cell Copper (Cu) Colorimetric Assay Kit (Elabscience Biotechnology, Wuhan, China) was selected to detect the copper ion content in SW480 cells, and the BCA Protein Colorimetric Assay Kit (Elabscience Biotechnology, Wuhan, China) was also used to determine the total protein concentration.

Techniques: Incubation, Western Blot, Expressing, Fluorescence

Journal: Heliyon

Article Title: A novel lnc-LAMC2-1:1 SNP promotes colon adenocarcinoma progression by targeting miR-216a-3p/HMGB3

doi: 10.1016/j.heliyon.2022.e12342

Figure Lengend Snippet: The lnc-LAMC2-1:1 SNP is a sponge of miR-216a-3p . Bar graphs of (a) and (b) show the relative luciferase activity of vectors containing SW480. A dual-luciferase reporter assay was performed, and the co-transfection of lnc-LAMC2-1:1 SNP and miR-216a-3p reduced the luciferase activity.

Article Snippet: The SW480 cell line was obtained from COSMOBIO Company in China and was grown in DMEM supplemented with 10% FBS (GIBCO) and 1% antibiotics (50 U/mL penicillin and 50 μg/mL streptomycin) in a humidified atmosphere of 5% CO2 at 37 °C.

Techniques: Luciferase, Activity Assay, Reporter Assay, Cotransfection

Journal: Heliyon

Article Title: A novel lnc-LAMC2-1:1 SNP promotes colon adenocarcinoma progression by targeting miR-216a-3p/HMGB3

doi: 10.1016/j.heliyon.2022.e12342

Figure Lengend Snippet: The lnc-LAMC2-1:1 SNP positively regulates HMGB3 by sponging miR-126-3p (a) mRNA expression of HMGB3 in SW480 cells. Cells were transfected with mimics of NC, miR-126-3p mimics, ASO-NC, and ASO-miR-126-3p. (b) mRNA expression of HMGB3 in SW480 cells. Cells were transfected with pcDNA3.1, lnc-LAMC2-1:1 SNP , and lnc-LAMC2-1:1-wt. (c) Complementary sequences of HMGB3 and miR-126-3p in the StarBase database. (d) A dual-luciferase reporter assay was performed, and the co-transfection of miR-126-3p and HMGB3 reduced the luciferase activity.

Article Snippet: The SW480 cell line was obtained from COSMOBIO Company in China and was grown in DMEM supplemented with 10% FBS (GIBCO) and 1% antibiotics (50 U/mL penicillin and 50 μg/mL streptomycin) in a humidified atmosphere of 5% CO2 at 37 °C.

Techniques: Expressing, Transfection, Luciferase, Reporter Assay, Cotransfection, Activity Assay

Journal: International Journal of Molecular Sciences

Article Title: Radiosensitization to γ-Ray by Functional Inhibition of APOBEC3G

doi: 10.3390/ijms23095069

Figure Lengend Snippet: The enhancement ratios at 10% survival (ER 10 ) to cell lines.

Article Snippet: A375 (purchased from ATCC), DU145 (purchased from RIKEN BRC, Tsukuba, Japan), MDA-MB-231 (purchased from ATCC), MIAPaCa2 (obtained from National Cancer Center), SAS (purchased from JCRB Cell Bank, Osaka, Japan), SW480 (purchased from JCRB Cell Bank), and U2OS (purchased from ATCC) cells were cultured in Dulbecco’s Modified Eagle’s Medium (Gibco).

Techniques: