Journal: bioRxiv

Article Title: MCC950/CRID3 potently targets the NACHT domain of wildtype NLRP3 but not disease-associated mutants for inflammasome inhibition

doi: 10.1101/634493

Figure Lengend Snippet: (A-F) Wild type (A- B), Nlrp3 L351P/L351P (C-D) or Nlrp3 A350V/A350V (E-F) BMDMs that were transduced with a Cre-recombinase expressing lentiviral vector, were left unstimulated (Ctrl) or stimulated with LPS and then left untreated or treated with ATP or nigericin (Nig) in absence or presence of MCC950/CRID3. Supernatants was analyzed for IL-1β (A,C,E) and IL-18 (B,D,F) secretion. Graphs show mean ± s.d. of triplicate wells and represent three independent experiments.

Article Snippet: To induce lentivirus production, the lentiviral GFP.Cre empty vector (20781, Addgene) together with 2 nd generation packaging vector psPAX2 (12260, Addgene) and VSV-G-expressing envelope plasmid pCMV-VSV-G (8454, Addgene) were transfected into HEK293 T cells using jetPRIME transfection reagent (114-15, PolyPlus Transfect).

Techniques: Transduction, Expressing, Plasmid Preparation

Journal: iScience

Article Title: The transcription factor IRF4 determines the anti-tumor immunity of CD8 + T cells

doi: 10.1016/j.isci.2023.108087

Figure Lengend Snippet: Key resources table

Article Snippet: The retroviral particles were generated by transfecting plat-E cells with the IRF4-GFP vector or the empty GFP-control vector, following the manufacturer's instructions (Cell Biolabs).

Techniques: Virus, Plasmid Preparation, Recombinant, Staining, Software

Journal: International Journal of Oral Science

Article Title: AFF4 regulates osteogenic differentiation of human dental follicle cells

doi: 10.1038/s41368-020-0083-9

Figure Lengend Snippet: Expression of AFF4 in DFCs. a Quantitative RT-PCR shows that the expression of AFF4 is the highest among AFF family members. n = 3. *** P < 0.001, and * P < 0.05. b Quantitative RT-PCR of AFF family members after osteogenic differentiation induction for 3 and 7 days. n = 3. *** P < 0.001, ** P < 0.01, and * P < 0.05. c Western blot analyses of AFF4 after osteogenic differentiation induction for 3 and 7 days. The quantitation of the WB is shown

Article Snippet: For overexpression experiments, lentiviruses expressing empty vectors GFP and AFF4 were purchased from Genechem (Shanghai, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Quantitation Assay

Journal: International Journal of Oral Science

Article Title: AFF4 regulates osteogenic differentiation of human dental follicle cells

doi: 10.1038/s41368-020-0083-9

Figure Lengend Snippet: Depletion of AFF4 decreases osteogenic differentiation of DFCs. a Quantitative RT-PCR showing the successful knockdown of AFF4 . n = 3. *** P < 0.001. b Western blot analysis and semiquantification of AFF4. c Representative ALP staining images of DFCs. d Quantitative assessment of ALP activity. n = 5. ** P < 0.01. e Representative images of ARS staining. f Quantitative assessment of calcium mineralization. n = 5. ** P < 0.01. g – j Quantitative RT-PCR demonstrating the reduced expression of DLX5 , SP7 and BGLAP . n = 3. * P < 0.05, and ** P < 0.01

Article Snippet: For overexpression experiments, lentiviruses expressing empty vectors GFP and AFF4 were purchased from Genechem (Shanghai, China).

Techniques: Quantitative RT-PCR, Knockdown, Western Blot, Staining, Activity Assay, Expressing

Journal: International Journal of Oral Science

Article Title: AFF4 regulates osteogenic differentiation of human dental follicle cells

doi: 10.1038/s41368-020-0083-9

Figure Lengend Snippet: Overexpression of AFF4 promotes osteogenic differentiation of DFCs. a Quantitative RT-PCR shows that the mRNA level of AFF4 is significantly increased. n = 3. *** P < 0.001. b Western blot analysis and semiquantification of AFF4. c Representative ALP staining images of DFCs. Increased expression of AFF4 enhances the intensity of the staining. d Quantitative ALP activity assessment. n = 5. *** P < 0.001. e Representative ARS staining images of DFCs. f Quantitative calcium mineralization assessment. n = 5. ** P < 0.01. g – j Quantitative RT-PCR displays the increased mRNA expression of DLX , SP7 and BGLAP . n = 3. * P < 0.05 and ** P < 0.01

Article Snippet: For overexpression experiments, lentiviruses expressing empty vectors GFP and AFF4 were purchased from Genechem (Shanghai, China).

Techniques: Over Expression, Quantitative RT-PCR, Western Blot, Staining, Expressing, Activity Assay

Journal: International Journal of Oral Science

Article Title: AFF4 regulates osteogenic differentiation of human dental follicle cells

doi: 10.1038/s41368-020-0083-9

Figure Lengend Snippet: AFF4 regulates the expression of ALKBH1. a , b Quantitative RT-PCR, western blot and quantification of ALKBH1 after the depletion of AFF4 . n = 3. ** P < 0.01. c , d Quantitative RT-PCR, western blot and quantification of ALKBH1 after AFF4 overexpression. n = 3. *** P < 0.001. e ChIP assay for AFF4 demonstrates that it binds to the promoter region of ALKBH1 . n = 4. ** P < 0.01

Article Snippet: For overexpression experiments, lentiviruses expressing empty vectors GFP and AFF4 were purchased from Genechem (Shanghai, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Over Expression

Journal: International Journal of Oral Science

Article Title: AFF4 regulates osteogenic differentiation of human dental follicle cells

doi: 10.1038/s41368-020-0083-9

Figure Lengend Snippet: Overexpression of ALKBH1 partially rescues osteogenic differentiation of AFF4 -deficient DFCs. a The mRNA level of ALKBH1 . n = 3. *** P < 0.001. b Western blot analyses of ALKBH1 and AFF4. Semi-quantifications of WB are shown. c , d Representative images of ALP staining and quantification of ALP activity. n = 5. * P < 0.05, and ** P < 0.01. e , f Representative images of ARS staining and quantitative analyses of calcium mineralization. n = 5. * P < 0.05, and ** P < 0.01

Article Snippet: For overexpression experiments, lentiviruses expressing empty vectors GFP and AFF4 were purchased from Genechem (Shanghai, China).

Techniques: Over Expression, Western Blot, Staining, Activity Assay

Journal: International Journal of Oral Science

Article Title: AFF4 regulates osteogenic differentiation of human dental follicle cells

doi: 10.1038/s41368-020-0083-9

Figure Lengend Snippet: The primer sequences for qRT-PCR and siRNA

Article Snippet: For overexpression experiments, lentiviruses expressing empty vectors GFP and AFF4 were purchased from Genechem (Shanghai, China).

Techniques: