Journal: PLOS ONE

Article Title: IGF2BP3 regulates the expression of RRM2 and promotes the progression of rheumatoid arthritis via RRM2/Akt/MMP-9 pathway

doi: 10.1371/journal.pone.0303593

Figure Lengend Snippet: A. The expression of RRM2 was detected via western blotting in synovial specimens from patients with RA and normal controls. The relative expression of RRM2 was shown in histogram. **p<0.01, compared to normal controls. B. MH7A cells were treated with TNF-α (10 ng/mL) and IL-1β (10 mg/L) for 24 h, and the expression of RRM2 was determined via western blotting. The relative expression of RRM2 was shown in histogram. **p<0.01, compared with untreated group. ##p<0.01, compared with untreated group. C. MH7A cells were treated with RRM2 shRNA lentivirus and lentiviral controls for 24 h. The relative expression of RRM2 was determined via western blotting and shown in histogram. **p<0.01, compared with control lentivirus group. D. MTT assay. MH7A cells were infected with RRM2 shRNA lentivirus and lentiviral controls for 24, 48, and 72 h, and cell viability was determined via MTT assay. **p<0.01. E. Clone formation assay. MH7A cells were infected with RRM2 shRNA and control shRNA lentivirus and cultured for 2 weeks. Colony formation assay was performed as described in Materials and Methods. The colony number was shown in histogram. **p<0.01, compared with control shRNA group. F. Migration and invasion assays. MH7A cells were infected with RRM2 shRNA and control shRNA lentiviruses and treated with 10 ng/mL of TNF-α. The migratory and invasive abilities were detected using transwell Boyden chamber coated without or with a Matrigel basement membrane matrix after 48 h. Original magnification ×100.

Article Snippet: RRM2 Lentiviral copy DNA Open Reading Frame Clone, Human, C-GFPSpark® tag (Cat: HG18284-ACGLN) and pLV-C-GFPSpark lentivirus control plasmid (Cat: LVCV-35) were purchased from SinoBiological corporation (Beijing, China).

Techniques: Expressing, Western Blot, shRNA, MTT Assay, Infection, Tube Formation Assay, Cell Culture, Colony Assay, Migration, Membrane

Journal: PLOS ONE

Article Title: IGF2BP3 regulates the expression of RRM2 and promotes the progression of rheumatoid arthritis via RRM2/Akt/MMP-9 pathway

doi: 10.1371/journal.pone.0303593

Figure Lengend Snippet: A. MH7A cells were treated with TNF-α (10 ng/mL) and IL-1β (10 mg/L) for 24 h, and the expression of RRM2 was determined via western blotting. B. The relative expression of IGF2BP3, RRM2, MMP-9 and MMP-1 was shown in histogram. **p<0.01, compared with control group. C. MH7A cells were infected with IGF2BP3 shRNA and control shRNA lentiviruses and treated with 10 ng/mL of TNF-α for 24 h. The expression of IGF2BP3, RRM2, MMP-9, and MMP-1 was detected via western blotting. D. The relative expression of IGF2BP3, RRM2, MMP-9 and MMP-1 was shown in histogram. **p<0.01, compared with control shRNA group.

Article Snippet: RRM2 Lentiviral copy DNA Open Reading Frame Clone, Human, C-GFPSpark® tag (Cat: HG18284-ACGLN) and pLV-C-GFPSpark lentivirus control plasmid (Cat: LVCV-35) were purchased from SinoBiological corporation (Beijing, China).

Techniques: Expressing, Western Blot, Infection, shRNA

Journal: PLOS ONE

Article Title: IGF2BP3 regulates the expression of RRM2 and promotes the progression of rheumatoid arthritis via RRM2/Akt/MMP-9 pathway

doi: 10.1371/journal.pone.0303593

Figure Lengend Snippet: A. MTT assay. MH7A cells were infected with IGF2BP3 shRNA lentivirus and control shRNA lentivirus was treated with TNF-α (10 ng/mL) for 24, 48, and 72 h. The cell viability was determined using MTT assay. **p<0.01, compared with control shRNA group. B. Clone formation assay. MH7A cells were infected with IGF2BP3 shRNA and control shRNA lentivirus and cultured for 2 weeks. The colony number was shown in histogram. **p<0.01, compared with control shRNA group. C. Migration and invasion assays. The effects of IGF2BP3 knockdown on invasion and migration were detected using transwell Boyden chamber coated with or without a Matrigel basement membrane matrix after 48 h. Original magnification ×100. D. MTT assay. MH7A cells were treated with IGF2BP3 shRNA lentivirus and overexpressed control lentivirus, control shRNA lentivirus and overexpressed RRM2 lentivirus, IGF2BP3 shRNA lentivirus and overexpressed control lentivirus, and IGF2BP3 shRNA lentivirus and overexpressed RRM2 lentivirus for 24, 48, and 72 h. Cell viability was determined using MTT assay. **p<0.01, compared with control group.

Article Snippet: RRM2 Lentiviral copy DNA Open Reading Frame Clone, Human, C-GFPSpark® tag (Cat: HG18284-ACGLN) and pLV-C-GFPSpark lentivirus control plasmid (Cat: LVCV-35) were purchased from SinoBiological corporation (Beijing, China).

Techniques: MTT Assay, Infection, shRNA, Tube Formation Assay, Cell Culture, Migration, Membrane

Journal: PLOS ONE

Article Title: IGF2BP3 regulates the expression of RRM2 and promotes the progression of rheumatoid arthritis via RRM2/Akt/MMP-9 pathway

doi: 10.1371/journal.pone.0303593

Figure Lengend Snippet: A. MH7A cells were infected with RRM2 shRNA or control shRNA lentiviruses for 48 h. The expression of RRM2, phosphorylated Akt, total Akt, and MMP-9 was detected via western blotting. The relative expression of RRM2, p-Akt, Akt and MMP-9 was shown in histogram. **p<0.01, compared with control shRNA group. B. The MH7A cells infected with RRM2 shRNA or control shRNA lentiviruses were treated with Akt inhibitor for 24 h. The expression of pAkt-S473, total Akt, and MMP-9 was detected via western blotting. The relative expression of RRM2, p-Akt, Akt, MMP-1 and MMP-9 was shown in histogram. **p<0.01, ##p<0.01, compared with control group.

Article Snippet: RRM2 Lentiviral copy DNA Open Reading Frame Clone, Human, C-GFPSpark® tag (Cat: HG18284-ACGLN) and pLV-C-GFPSpark lentivirus control plasmid (Cat: LVCV-35) were purchased from SinoBiological corporation (Beijing, China).

Techniques: Infection, shRNA, Expressing, Western Blot

Journal: Cell Death & Disease

Article Title: MicroRNA-184 promotes apoptosis of trophoblast cells via targeting WIG1 and induces early spontaneous abortion

doi: 10.1038/s41419-019-1443-2

Figure Lengend Snippet: a HTR8 cells were infected with negative control lentivirus and miR-184 overexpression lentivirus for 72 h, and GFP-positive cells were sorted out. Flow cytometry was used to detect the infection efficiency. b We also used RT-PCR to detect the mRNA expression of miR-184 in NC and miR-184 cells. Unpaired student’s t -test (two-tailed) was used for comparison between two groups. **** P < 0.0001. Data represent mean ± SEM. c CCK8 was used to detect cell proliferation of NC and miR-184 cells, which indicated that miR-184 could markedly inhibit cell proliferation. Unpaired student’s t -test (two-tailed) was used for comparison between two groups. ** P < 0.01. Data represent mean ± SEM. d After culturing NC and miR-184 cells for 48 h, the Annexin V staining result showed that miR-184 could increase cell apoptosis significantly. Unpaired student’s t -test (two-tailed) was used for comparison between two groups. * P < 0.05. Data represent mean ± SEM. e We examined the invasiveness of NC and miR-184 cells via transwell assay, finding that miR-184 had little effect on the invasiveness of trophoblast cells. Scale bar: 100 μm. Unpaired student’s t -test (two-tailed) was used for comparison between two groups. Data represent mean ± SEM. These experiments were repeated three times. (miR-184: HTR8 cells infected with miR-184 overexpression lentivirus; NC: HTR8 cells infected with negative control lentivirus)

Article Snippet: Protein levels were detected after 48 h. To generate hsa-miR-184 overexpression cells, HTR8 cell line was transfected by lentivirus encoding hsa-miR-184 and its control lentivirus purchased from Suzhou GenePharma.

Techniques: Infection, Negative Control, Over Expression, Flow Cytometry, Reverse Transcription Polymerase Chain Reaction, Expressing, Two Tailed Test, Staining, Transwell Assay

Journal: Cell Death & Disease

Article Title: MicroRNA-184 promotes apoptosis of trophoblast cells via targeting WIG1 and induces early spontaneous abortion

doi: 10.1038/s41419-019-1443-2

Figure Lengend Snippet: a We detected some molecules related to the poptotic pathway by western blot analysis, including caspase-9, -8, -3, -7, Fas, and FasL, and only the expression of Fas was changed after miR-184 overexpression. b Flow cytometry was used to examine the expression of Fas on NC and miR-184 cells, showing that the percentage of Fas-positive cells was increased in miR-184 cells, as well as the mean fluorescence intensity. Unpaired student’s t -test (two-tailed) was used for comparison between two groups. **** P < 0.0001. Data represent mean ± SEM. c Meanwhile, RT-PCR was applied to detect the mRNA level of Fas in two groups. Unpaired student’s t -test (two-tailed) was used for comparison between two groups. ** P < 0.01. Data represent mean ± SEM. d After stimulation with various concentrations of anti-Fas activating antibody (zero, 1, 3, 5 μg /mL) for 24 h, the apoptosis rate of NC and miR-184 cells was detected. Two-way ANOVA was used for comparison between these groups. ** P < 0.01, *** P < 0.001, **** P < 0.0001. Data represent mean ± SEM. e The protein level of caspase-3, -8, and cleaved caspase-3, -8 in NC and miR-184 cells after stimulation with different concentrations of anti-Fas activating antibody (zero, 3, 5 μg /mL) was also examined by western blot analysis. These experiments were repeated three times. (miR-184: HTR8 cells infected with miR-184 overexpression lentivirus; NC: HTR8 cells infected with negative control lentivirus.)

Article Snippet: Protein levels were detected after 48 h. To generate hsa-miR-184 overexpression cells, HTR8 cell line was transfected by lentivirus encoding hsa-miR-184 and its control lentivirus purchased from Suzhou GenePharma.

Techniques: Western Blot, Expressing, Over Expression, Flow Cytometry, Fluorescence, Two Tailed Test, Reverse Transcription Polymerase Chain Reaction, Infection, Negative Control

Journal: Cell reports

Article Title: Characterization of a small molecule inhibitor of disulfide reductases that induces oxidative stress and lethality in lung cancer cells

doi: 10.1016/j.celrep.2022.110343

Figure Lengend Snippet: Key resources table

Article Snippet: Pathway control lentivirus , GenTarget , Path-Ctr5-PBS.

Techniques: Virus, Control, Recombinant, Purification, Lysis, Saline, Staining, BIA-KA, Annexin V Assay, Multiplex sample analysis, Knock-Out, Software, Membrane, Imaging, Flow Cytometry, Mass Spectrometry

Journal: iScience

Article Title: FOXO1 stimulates tip cell-enriched gene expression in endothelial cells

doi: 10.1016/j.isci.2024.109161

Figure Lengend Snippet:

Article Snippet: EGFP control lentivirus , VectorBuilder , VB010000.

Techniques: Virus, CRISPR, Control, Recombinant, Modification, Saline, Chromatin Immunoprecipitation, Library Quantification, Biomarker Discovery, Selection, Plasmid Preparation, Software