Journal: Frontiers in Pharmacology

Article Title: DNA Methylation of PTGIS Enhances Hepatic Stellate Cells Activation and Liver Fibrogenesis

doi: 10.3389/fphar.2018.00553

Figure Lengend Snippet: The siRNA sequences used in RNA interference analysis.

Article Snippet: Small interfering RNA (siRNA) oligonucleotides used for knockdown gene and a negative scrambled siRNA was used in parallel were synthesized by GenePharma (Shanghai, China).

Techniques: Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: miR-30 decreases multidrug resistance in human gastric cancer cells by modulating cell autophagy

doi: 10.3892/etm.2017.5354

Figure Lengend Snippet: Quantification of miR-30a expression in different cell groups by RT-PCR analysis. miR-30a expression was decreased in chemoresistant SGC-7901/CDDP cells compared with chemosensitive SGC-7901 cells. When transfected with miR-30a mimics or NC miRNA, miR-30a expression in the SGC7901/CDDP cells was notably increased or decreased, respectively. **P<0.01 vs. SGC7901; ***P<0.001 vs. SGC7901/CDDP. CDDP, cisplatin.

Article Snippet: miR-30a mimics, and scramble miRNA negative control (NC miRNA) were synthesized by Shanghai GenePharma Co., Ltd. (Shanghai, China).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Transfection

Journal: Experimental and Therapeutic Medicine

Article Title: miR-30 decreases multidrug resistance in human gastric cancer cells by modulating cell autophagy

doi: 10.3892/etm.2017.5354

Figure Lengend Snippet: Expression analysis of P-gp and MDR1 in different cell groups. Total protein or RNA was extracted for P-gp and MDR1 analysis. (A) The protein levels of P-gp in SGC7901 and SGC7901/CDDP cells were determined by western blot analysis. (B) P-gp expression was decreased in SGC7901/CDDP cells transfected with miR-30a mimics. (C) The mRNA levels of MDR1 were detected by real time PCR analysis. MDR1 mRNA expression was decreased in SGC7901 cells transfected with miR-30a mimics. Control, SGC7901/CDDP cells without transfection treatment; NC miRNA, SGC7901/CDDP cells transfected with NC miRNA; miR-30a mimics, SGC7901/CDDP cells transfected with miR-30a mimics. Data were expressed as the means ± SD of three independent experiments. ***P<0.001 vs. SGC7901 group; ###P<0.001 vs. SGC7901/CDDP control cells; **P<0.01 vs. SGC7901/CDDP control group. P-gp, P-glycoprotein; CDDP, cisplatin.

Article Snippet: miR-30a mimics, and scramble miRNA negative control (NC miRNA) were synthesized by Shanghai GenePharma Co., Ltd. (Shanghai, China).

Techniques: Expressing, Western Blot, Transfection, Real-time Polymerase Chain Reaction, Control

Journal: Experimental and Therapeutic Medicine

Article Title: miR-30 decreases multidrug resistance in human gastric cancer cells by modulating cell autophagy

doi: 10.3892/etm.2017.5354

Figure Lengend Snippet: Effects of miR-30a on cell apoptosis and LC3-II/LC3-I expression. (A) Flow cytometric analysis of apoptotic SGC-7901 cells or SGC7901/CDDP cells (control SGC7901/CDDP cells without transfection and SGC7901/CDDP cells transfected with NC siRNA or miR-30a mimics) 24 h after treatment with 5 µg/ml CDDP. (B) Western blot analysis of LC3-II/LC3-I expression. (C) Relative LC3-II/LC3-I expression determined from density analysis. Data were expressed as the means ± SD of three independent experiments. Control, SGC7901/CDDP control cells without transfection treatment; NC miRNA, SGC7901/CDDP cells transfected with NC miRNA; miR-30a mimics, SGC7901/CDDP cells transfected with miR-30a mimics. ***P<0.001 vs. SGC7901 group; ###P<0.001 vs. SGC7901/CDDP control cells. LC, light chain; CDDP, cisplatin.

Article Snippet: miR-30a mimics, and scramble miRNA negative control (NC miRNA) were synthesized by Shanghai GenePharma Co., Ltd. (Shanghai, China).

Techniques: Expressing, Control, Transfection, Western Blot

Journal: Oncology Reports

Article Title: Ginsenoside compound K exerts antitumour effects in renal cell carcinoma via regulation of ROS and lncRNA THOR

doi: 10.3892/or.2021.7989

Figure Lengend Snippet: Silencing of THOR inhibits the tumorigenesis of renal cell carcinoma cells. (A) Cells were transfected as indicated for 24 h, and the expression of THOR was measured. (B) Cell viability was measured via an MTT assay after transfection. (C) Migration and invasion were measured after transfection. (D) Cellular apoptosis was measured after transfection. (E) Cellular lysates were subjected to western blotting analysis with the indicated antibodies after transfection. (F) Cell cycle distribution was analysed after transfection. The data are presented as the mean ± SD of at least three independent experiments. **P<0.01 and ***P<0.001 vs. control group. si-, small interfering RNA; NC, negative control; THOR, testis associated oncogenic.

Article Snippet: Small interfering RNA (siRNA) targeting testis associated oncogenic (THOR) (si-THOR), negative control scramble siRNA (si-NC), pcDNA3.1 expressing THOR and empty control were synthesized and obtained from Suzhou GenePharma Co., Ltd. Transfections were performed using Lipofectamine ® 2000 (Thermo Fisher Scientific, Inc.), according to the manufacturer's protocols.

Techniques: Transfection, Expressing, MTT Assay, Migration, Western Blot, Control, Small Interfering RNA, Negative Control

Journal: Journal of Clinical & Cellular Immunology

Article Title: MicroRNA Regulation of Proinflammatory Response in X-linked Adrenoleukodystrophy

doi: 10.4172/2155-9899.1000349

Figure Lengend Snippet: Figure 3: miR-323-5p mediates AMPKα1 deletion-induced iNOS expression in Abcd1-KO mice mixed glial cells. (A) Abcd1-KO mixed glial cells were silenced for scrambled control (Scr) or AMPKα1 as described in section 2. iNOS mRNA expression (B) was induced and miR-323-5p expression decreased (C) in Abcd1-KO mixed glial cells deleted for AMPKα1 using lentiviral particles. Abcd1-KO mixed glial cells deleted or AMPKα1 and co-transfected with miR-323-5p mimic (MIM) or inhibitors (INB) modulated iNOS mRNA (D) and protein (E) levels. Mimic and inhibitor transfected cells were compared with negative control (NGC) transfected cells. Results represent the mean ± SD of triplicates from three different experiments. ***p<0.001.

Article Snippet: Lentiviral vector mediated knockdown of AMPKα1 in Abcd1-KO mice mixed glial cells Transduction-ready mouse shRNA lentiviral particles (106 TU/ml) for AMPKα1 (consisting of a pool of 3-5 constructs and puromycin selection gene; sc-29674-V) and control scrambled shRNA lentiviral particles (Scr) (106 TU/ml, sc-108080) were purchased from Santa Cruz Biotechnology (Dallas, TX).

Techniques: Expressing, Control, Transfection, Negative Control

Journal: Animal Models and Experimental Medicine

Article Title: The sigma‐1 receptor‐TAMM41 axis modulates neuroinflammation and attenuates memory impairment during the latent period of epileptogenesis

doi: 10.1002/ame2.12341

Figure Lengend Snippet: Knockdown of TAMM41 (translocator assembly and maintenance 41 homolog) reversed the anti‐inflammatory and memory‐improving action of PRE‐084. (A) Experimental schematic showing epilepsy induction, drug treatment, siRNA (small interfering RNA) delivery, and testing processes. (B) Western blot image indicating that protein expression was knocked down by TAMM41 siRNA ( n = 6 in each group). (C) Summarized results from (B). (D–F) mRNA (messenger RNA) levels of inflammatory mediators in the hippocampus. (G–I) Recognition indices in the OLT, NORT, and PST. Epilepsy was induced by kainic acid injection. PRE‐084 (5 mg kg −1 ) was administered intraperitoneally the day after the inducement of epilepsy. Behavior tests were performed from the fifth to the eighth day ( n = 10 in each group). mRNA expression levels of TNF‐α, IL‐1β, and iNOS were quantified using qPCR (quantitative polymerase chain reaction) on the eighth day ( n = 6 in each group). Data were expressed as mean ± SEM (standard error of the mean) and analyzed using Student's t ‐test. * p < 0.05; ** p < 0.01; *** p < 0.001, and **** p < 0.0001. IL‐1β, interleukin‐1β; iNOS, inducible nitric oxide synthase; NORT, novel object recognition test; OLT, object location test; PST, pattern separation test; TNF‐α, tumor necrosis‐α.

Article Snippet: TAMM41 small interfering RNA (siRNA, Tamm41‐585: 5′‐CCUCAUGCUACCUGAAAGCUUTT‐3′, 5′‐AAGCUUUCAGGUAGCAUGAGGTT‐3′) and the corresponding scramble siRNA (as the negative control [NC]) were obtained from Santa Cruz Biotechnology (Shanghai) Co., Ltd. Adeno‐associated viruses containing the TAMM41 coding sequence (pHBAAV‐CMV‐MCS‐3flag‐T2A‐ZsGreen, abbreviation: AAV‐TAMM41) and the vector virus (AAV‐CON) were obtained from Hanbio Company.

Techniques: Knockdown, Small Interfering RNA, Western Blot, Expressing, Injection, Real-time Polymerase Chain Reaction

Journal: Animal Models and Experimental Medicine

Article Title: The sigma‐1 receptor‐TAMM41 axis modulates neuroinflammation and attenuates memory impairment during the latent period of epileptogenesis

doi: 10.1002/ame2.12341

Figure Lengend Snippet: Knockdown of TAMM41 (translocator assembly and maintenance 41 homolog) reversed the gliosis‐inhibition and neuron‐protective actions of PRE‐084. (A) Typical images of GFAP + astrocytes. (B) Typical images of Iba1 + microglia. (C) Typical images of MAP‐2 + neurons. (D) Quantification of GFAP + astrocytes in the hippocampal, DG, and CA1 regions. (E) Quantification of microglia in the hippocampal, DG, and CA1 regions. (F) Quantification of MAP‐2 + neurons in the total hippocampal, DG, and CA1 regions. The representative characteristics are marked with arrows. Epilepsy was induced by kainic acid injection, and the mice were killed on the eighth day. PRE‐084 (5 mg kg −1 ) was administered intraperitoneally the day after the inducement of epilepsy. DG, dentate gyrus; GFAP, glial fibrillary acidic protein; Iba1, ionized calcium binding adapter; MAP‐2, microtubule‐associated protein‐2; NC‐siRNA, negative control siRNA. Data were expressed as mean ± SEM (standard error of the mean) and analyzed using Student's t ‐test. * p < 0.05; ** p < 0.01; and *** p < 0.001. Each group comprised three mice.

Article Snippet: TAMM41 small interfering RNA (siRNA, Tamm41‐585: 5′‐CCUCAUGCUACCUGAAAGCUUTT‐3′, 5′‐AAGCUUUCAGGUAGCAUGAGGTT‐3′) and the corresponding scramble siRNA (as the negative control [NC]) were obtained from Santa Cruz Biotechnology (Shanghai) Co., Ltd. Adeno‐associated viruses containing the TAMM41 coding sequence (pHBAAV‐CMV‐MCS‐3flag‐T2A‐ZsGreen, abbreviation: AAV‐TAMM41) and the vector virus (AAV‐CON) were obtained from Hanbio Company.

Techniques: Knockdown, Inhibition, Injection, Binding Assay, Negative Control

Journal:

Article Title: gp-91 mediates histone deacetylase inhibition-induced cardioprotection

doi: 10.1016/j.bbamcr.2010.04.007

Figure Lengend Snippet: Western blot analysis of gp-91 in H9c2 cardiomyoblasts transfected with or without gp-91 siRNA. (A): H9c2 cardiomyoblast were transfected with gp-91 (500 nmol/L) or scrambled siRNA (500 nmol/L) for 24 h to prepare protein lysates and following immunoblotting assay. (B) The LDH release of H9c2 myocytes in culture medium at the end of reperfusion (n=4/group). (C): Cell viability was measured using the MTT cell viability assay (n=4/group). H9c2 cardiomyoblast cells were exposed to simulated ischemia (KCN and 2-deoxy-D-glucose) and description of simulated ischemia is provided under METHODS. HDAC inhibitor, trichostatin A (50 nmol/L) was maintained in culture medium. Gp-91 and negative control siRNA transfections are described as under METHODS. SI: simulated ischemia. LDH: lactate dehydrogenase. Results are expressed as means ± SE and the values expressed as percentages of control values. *P < 0.05.

Article Snippet: Gp-91 polyclonal rabbit antibody, β-actin, gp-91 siRNA and negative control scrambled siRNA were purchased from Santa Cruz Biotechnology Inc.

Techniques: Western Blot, Transfection, Viability Assay, Negative Control, Control