Journal: Bioengineered

Article Title: Circular RNA circ-LARP1B contributes to cutaneous squamous cell carcinoma progression by targeting microRNA-515-5p/TPX2 microtubule nucleation factor axis

doi: 10.1080/21655979.2021.2019172

Figure Lengend Snippet: miR-515-5p overexpression-mediated anti-tumor effects in CSCC cells are overturned by the accumulation of TPX2. (a-l) CSCC cells were transfected with miR-NC, miR-515-5p, miR-515-5p + pcDNA or miR-515-5p + TPX2. (a and b) The mRNA and protein expression of TPX2 was examined in CSCC cells by RT-qPCR and Western blot assay. (c) Cell viability was analyzed by MTT assay. (d) The colony formation ability was analyzed by colony formation assay. (e) Cell apoptosis rate was analyzed by flow cytometry. (f and g) Cell migration and invasion abilities were analyzed by wound healing assay and transwell invasion assay. (h) The percentages of CSCC cells in G0/G1 phase, S phase, and G2/M phase were analyzed by flow cytometry. (i-k) Cell glycolytic metabolism was analyzed using Glucose Uptake Colorimetric Assay kit, Lactate Assay Kit II, and ATP colorimetric Assay kit. (l) Western blot assay was conducted to analyze the protein level of HK-2 in CSCC cells. * P < 0.05, ** P < 0.01.

Article Snippet: Small interfering (si)RNAs targeting circ-LARP1B (si-circ-LARP1B#1 and #2), siRNA negative control (si-NC), circ-LARP1B ectopic expression plasmid (circ-LARP1B), pLCDH-cir empty vector (pLCDH-cir), short hairpin (sh)RNA targeting circ-LARP1B (sh-circ-LARP1B), shRNA NC (sh-NC), miR-515-5p mimics (miR-515-5p), miR-NC, miR-515-5p inhibitor (anti-miR-515-5p), anti-miR-NC, TPX2 ectopic expression plasmid (TPX2), and pcDNA empty vector (pcDNA) were purchased from Ribobio (Shanghai, China) and GenePharma (Shanghai, China).

Techniques: Over Expression, Transfection, Expressing, Quantitative RT-PCR, Western Blot, MTT Assay, Colony Assay, Flow Cytometry, Migration, Wound Healing Assay, Transwell Invasion Assay, Colorimetric Assay, Lactate Assay

Journal: Molecular Medicine Reports

Article Title: circSAMD4A participates in the apoptosis and autophagy of dopaminergic neurons via the miR-29c-3p-mediated AMPK/mTOR pathway in Parkinson's disease

doi: 10.3892/mmr.2021.12179

Figure Lengend Snippet: circSAMD4A targets and negatively regulates miR-29c-3p. (A) Upper panel, TargetScan analysis showing the putative recognition sequence of miR-29c-3p in circSAMD4A. Lower panel, a dual-luciferase reporter was used to verify the interaction between circSAMD4A and miR-29c-3p in 293 cells. The relative expression levels of (B) circSAMD4A and (C) miR-29c-3p were detected in SH-SY5Y cells transfected with sh-circ, pcDNA-circSAMD4A or matched controls via reverse transcription-quantitative PCR. **P<0.01, ***P<0.001. sh-circ, sh-circSAMD4A; miR, microRNA; circSAMD4A, circular RNA sterile α motif domain containing 4A; WT, wild-type; MUT, mutant; sh, short hairpin RNA; NC, negative control.

Article Snippet: miR-29c-3p mimics (including miR control, cat. no. HMI0439, concentration: ≥10 6 TU/ml), miR-29c-3p inhibitor (including miR-29c-3p inhibitor NC, cat. no. HSTUD0440, concentration: ≥10 6 TU/ml) were purchased from Sigma-Aldrich (Shanghai) Trading Co., Ltd., shNC (cat. no. C01001, concentration: 10 8 TU/ml), sh-circ (cat. no. C03002, concentration: 10 8 TU/ml), pcDNA empty vector (cat. no. G01001, concentration: 10 8 TU/ml), and pcDNA-circSAMD4A (cat. no. G04002, concentration: 10 8 TU/ml) were purchased from Shanghai GenePharma Co., Ltd.

Techniques: Sequencing, Luciferase, Expressing, Transfection, Real-time Polymerase Chain Reaction, Mutagenesis, shRNA, Negative Control