control empty vectors nc  (Genechem)

Journal: Cell Biology and Toxicology

Article Title: Endothelial progenitor cell derived extracellular vesicles promotes wound healing in diabetic mice via activating mobilization and neovascularization

doi: 10.1007/s10565-025-10134-3

Figure Lengend Snippet: Overexpression of SNHG1 or HDAC6 partially reverses the pro-angiogenic effect of EPC-EVs on MAECs. MAECs were transfected with SNHG1 overexpression vectors (oe-SNHG1), HDAC6 overexpression vectors (oe-HDAC6), or empty vectors (oe-NC, control) in HG (30 mM) medium. A : Transfection efficiency of oe-SNHG1 and oe-HDAC6 was verified by qRT-PCR (n = 3). Cells were then co-treated with EPC-EVs-miR in HG medium. B : SNHG1 and HDAC6 expression was detected by qRT-PCR (n = 3). C : Expression of HDAC6, CD31, and VEGFA was measured by Western blot (n = 3). D : Cell viability was assessed by CCK-8 assay (n = 3). E : Angiogenic capacity was evaluated by tube formation assay (n = 3). F : Cell migration was measured by scratch wound healing assay (n = 3). Data are presented as mean ± standard deviation. Data in panel A were analyze using student’s t-test. Data in panels B , D , E , and F were analyzed using one-way ANOVA. Data in panel C were analyzed using two-way ANOVA, followed by Tukey’s multiple comparisons test. * p < 0.05 and ** p < 0.01

Article Snippet: For lentiviral infection, GFP-labeled miR-204-5p lentiviral overexpression vectors, SNHG1 lentiviral overexpression vectors, or control empty vectors (NC) (Genechem, Shanghai, China) were applied to transfection with 293 T cells (CRL-1573; ATCC, Manassas, VA, USA).

Techniques: Over Expression, Transfection, Control, Quantitative RT-PCR, Expressing, Western Blot, CCK-8 Assay, Tube Formation Assay, Migration, Wound Healing Assay, Standard Deviation

Journal: Heliyon

Article Title: CircACTR2 promotes bladder cancer progression through IKBKB-mediated NF-κB signaling pathway activation

doi: 10.1016/j.heliyon.2024.e30778

Figure Lengend Snippet: Sequences used in transfections.

Article Snippet: For IKBKB overexpression, the entire sequence of IKBKB was sub-cloned into the pcDNA3.1 vector (Invitrogen, Carlsbad, CA, USA) to generate the pcDNA3.1/IKBKB expression vector; the empty pcDNA3.1 vector was used as the negative control (NC).

Techniques: Transfection

Journal: Heliyon

Article Title: CircACTR2 promotes bladder cancer progression through IKBKB-mediated NF-κB signaling pathway activation

doi: 10.1016/j.heliyon.2024.e30778

Figure Lengend Snippet: IKBKB is involved in circACTR2-mediated BCa cell proliferation, invasion and migration. J82 cells were transfected with si-NC, si-circACTR2-1, or co-transfected with si-circACTR2-1 and pcDNA3.1/IKBKB for functional rescue assays. A-B. The proliferation ability of three groups of J82 cells was evaluated by CCK-8 assay (A) and EdU assay (B), respectively. C-D. The number of invaded or migrated cells was calculated using transwell invasion (C) and migration (D) assays after treatment with three different transfections. E. A wound healing assay was carried out in three groups of J82 cells to detect migration ability. **P < 0.01.

Article Snippet: For IKBKB overexpression, the entire sequence of IKBKB was sub-cloned into the pcDNA3.1 vector (Invitrogen, Carlsbad, CA, USA) to generate the pcDNA3.1/IKBKB expression vector; the empty pcDNA3.1 vector was used as the negative control (NC).

Techniques: Migration, Transfection, Functional Assay, CCK-8 Assay, EdU Assay, Wound Healing Assay

Journal: Aging (Albany NY)

Article Title: Metformin inhibits ovarian granular cell pyroptosis through the miR-670-3p/NOX2/ROS pathway

doi: 10.18632/aging.204745

Figure Lengend Snippet: Effects of NOX2 overexpression on the inhibitory effects of metformin on inflammation and KGN cell pyroptosis. ( A – C ) Cells were pretreated with or without 10 μM LPS for 24 h and then incubated with 20 μM metformin for another 12 h. ( A ) NOX activity was evaluated using a commercial NOX detection kit; ( B ) Quantification of NOX2 mRNA expression using RT-PCR; ( C ) Analysis of NOX2 protein levels using Western blotting; ( D ) Cells were transduced with PBS, LV-NC, or LV-NOX2 for 48 h. The transfection efficiency was evaluated using Western blot analysis; ( E – J ) Cells were pretreated with 10 μM LPS for 24 h or transfected with LV-NOX2 for 48 h and then incubated with 20 μM metformin for another 12 h; ( E ) Intracellular levels of ROS were measured using commercial kits; ( F ) The mRNA levels of NLRP3, ASC, caspase-1 and GSDMD were detected by RT-PCR; ( G ) The protein levels of NLRP3, pro-caspase-1, cleaved-caspase-1, ASC and GSDMD-N were detected by Western blot; ( H ) Levels of LDH in the cell culture medium were examined using an LDH cytotoxicity detection kit; ( I ) Caspase-1 activity was assessed using a colorimetric caspase-1 activity assay kit; ( J ) The levels of IL-1β, IL-18, IL-6, and TNF-α in the cell culture supernatant were determined by ELISA. Data are represented as the means ± SD from three independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns. not significant.

Article Snippet: The lentiviral vector encoding NOX2 (LV-NOX2) and a control empty lentiviral vector (LV-NC) were purchased from OriGene Technologies (Rockville, MD, USA).

Techniques: Over Expression, Incubation, Activity Assay, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Transduction, Transfection, Cell Culture, Enzyme-linked Immunosorbent Assay