Journal: Bioengineered

Article Title: Long non-coding RNA cancer susceptibility candidate 2 regulates the function of human fibroblast-like synoviocytes via the microRNA-18a-5p/B-cell translocation gene 3 signaling axis in rheumatoid arthritis.

doi: 10.1080/21655979.2021.2022075

Figure Lengend Snippet: Figure 5. BTG3 directly targets miR-18a-5p. (a) A schematic of the miR-18a-5p binding site in lncRNA CASC2 3’-UTR. (b) Binding between miR-18a-5p and BTG3 was confirmed using dual-luciferase reporter analysis. (c) mRNA expression of BTG3 in the sera of patients with RA and healthy volunteers. **P < 0.01 vs. mimic control; ##P < 0.01 vs. healthy control. miR, microRNA; lncRNA CASC2, long non-coding RNA cancer susceptibility candidate 2; RA, rheumatoid arthritis; BTG3, B-cell translocation gene 3. Experiments were repeated for three times.

Article Snippet: The control plasmid, lncRNA CASC2 plasmid, mimic control (cat. no. miR1N0000001-1-5; Guangzhou RiboBio Co., Ltd., Guangzhou, China), miR-18a-5p mimic (cat. no. miR10000072-1-5; Guangzhou RiboBio Co., Ltd., Guangzhou, China), inhibitor control (cat. no. miR2N0000001-1-5; Guangzhou RiboBio Co., Ltd., Guangzhou, China), miR-18a-5p inhibitor (cat. no. miR20000072-1-5; Guangzhou RiboBio Co., Ltd., Guangzhou, China), control small interfering (si) RNA, and BTG3 siRNA (Santa Cruz Biotechnology, Inc.) were transfected into HFLSs using Lipofectamine 2000 (Invitrogen; Thermo Fisher Scientific, Inc.) for 48 h referring to the manufacturer’s protocol.

Techniques: Binding Assay, Luciferase, Expressing, Control, Translocation Assay

Journal: Bioengineered

Article Title: Long non-coding RNA cancer susceptibility candidate 2 regulates the function of human fibroblast-like synoviocytes via the microRNA-18a-5p/B-cell translocation gene 3 signaling axis in rheumatoid arthritis.

doi: 10.1080/21655979.2021.2022075

Figure Lengend Snippet: Figure 6. BTG3 siRNA abolishes the influences of the miR-18a-5p inhibitor on HFLSs proliferation and apoptosis. The inhibitor control, miR-18a-5p inhibitor, control siRNA, and BTG3 siRNA were transfected into HFLSs for 48 h. (a) miR-18a-5p expression was evaluated in HFLSs. (b) RT-qPCR analysis of BTG3 in control siRNA- or BTG3 siRNA-transfected HFLSs. Evaluation of BTG3 (c) mRNA and (d) protein levels in transfected HFLSs using RT-qPCR and Western blotting. (e) HFLS viability was measured using an MTT assay. (f) Flow cytometric analysis of apoptotic cells in HFLSs. (g) Quantification of apoptotic cells. (h) Determination of Bax and Bcl-2 protein expression levels using Western blotting. qRT-PCR analysis of Bax (i) and Bcl-2 (j) mRNA levels in different groups. **P < 0.01 vs. inhibitor control; ##P < 0.01 vs. control siRNA; &&P < 0.01 vs. miR-18a-5p inhibitor + control siRNA. miR, microRNA; lncRNA CASC2, long non-coding RNA cancer susceptibility candidate 2; RA, rheumatoid arthritis; BTG3, B-cell translocation gene 3; RT-qPCR, reverse transcription-quantitative PCR; HFLSs, human fibroblast-like synoviocytes; siRNA, small interfering RNA. Experiments were repeated for three times.

Article Snippet: The control plasmid, lncRNA CASC2 plasmid, mimic control (cat. no. miR1N0000001-1-5; Guangzhou RiboBio Co., Ltd., Guangzhou, China), miR-18a-5p mimic (cat. no. miR10000072-1-5; Guangzhou RiboBio Co., Ltd., Guangzhou, China), inhibitor control (cat. no. miR2N0000001-1-5; Guangzhou RiboBio Co., Ltd., Guangzhou, China), miR-18a-5p inhibitor (cat. no. miR20000072-1-5; Guangzhou RiboBio Co., Ltd., Guangzhou, China), control small interfering (si) RNA, and BTG3 siRNA (Santa Cruz Biotechnology, Inc.) were transfected into HFLSs using Lipofectamine 2000 (Invitrogen; Thermo Fisher Scientific, Inc.) for 48 h referring to the manufacturer’s protocol.

Techniques: Control, Transfection, Expressing, Quantitative RT-PCR, Western Blot, MTT Assay, Translocation Assay, Reverse Transcription, Real-time Polymerase Chain Reaction, Small Interfering RNA

Journal: Bioengineered

Article Title: Long non-coding RNA cancer susceptibility candidate 2 regulates the function of human fibroblast-like synoviocytes via the microRNA-18a-5p/B-cell translocation gene 3 signaling axis in rheumatoid arthritis.

doi: 10.1080/21655979.2021.2022075

Figure Lengend Snippet: Figure 7. BTG3 siRNA transfection abolishes the miR-18a-5p inhibitor influences on inflammatory cytokine release in HFLSs. After transfection, the secretion of inflammatory cytokines, such as (a) TNF-a, (b) IL-1β, and (c) IL-6, and MMPs including MMP1 (d) and MMP3 (e) were determined via ELISAs in different groups. **P < 0.01 vs. inhibitor control; ##P < 0.01 vs. miR-18a-5p inhibitor + control siRNA. miR, microRNA; BTG3, B-cell translocation gene 3. Experiments were repeated for three times.

Article Snippet: The control plasmid, lncRNA CASC2 plasmid, mimic control (cat. no. miR1N0000001-1-5; Guangzhou RiboBio Co., Ltd., Guangzhou, China), miR-18a-5p mimic (cat. no. miR10000072-1-5; Guangzhou RiboBio Co., Ltd., Guangzhou, China), inhibitor control (cat. no. miR2N0000001-1-5; Guangzhou RiboBio Co., Ltd., Guangzhou, China), miR-18a-5p inhibitor (cat. no. miR20000072-1-5; Guangzhou RiboBio Co., Ltd., Guangzhou, China), control small interfering (si) RNA, and BTG3 siRNA (Santa Cruz Biotechnology, Inc.) were transfected into HFLSs using Lipofectamine 2000 (Invitrogen; Thermo Fisher Scientific, Inc.) for 48 h referring to the manufacturer’s protocol.

Techniques: Transfection, Control, Translocation Assay