Journal: Cell Reports Medicine

Article Title: Boosting NAD preferentially blunts Th17 inflammation via arginine biosynthesis and redox control in healthy and psoriasis subjects

doi: 10.1016/j.xcrm.2023.101157

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-CD38 , Cell Signaling , Cat# 92457; RRID: N/A.

Techniques: Flow Cytometry, Virus, Recombinant, Staining, Cell Stimulation, CyQUANT Assay, Proliferation Assay, Bicinchoninic Acid Protein Assay, In Vitro, Antioxidant Assay, Detection Assay, GSH Assay, Bioassay, Transcription Factor Assay, Extraction, Enzyme-linked Immunosorbent Assay, Control, Software, Cell Isolation, Transfection

Journal: Molecular Cell

Article Title: The IMiD target CRBN determines HSP90 activity toward transmembrane proteins essential in multiple myeloma

doi: 10.1016/j.molcel.2020.12.046

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-CD38 , Abcam , Cat.#: ab125038 RRID: AB_10976505 , .

Techniques: Generated, Isolation, Recombinant, Transfection, Clone Assay, shRNA, Software

Journal: Cancer research

Article Title: Targeting of CD38 by the tumor suppressor miR-26a serves as a novel potential therapeutic agent in multiple myeloma

doi: 10.1158/0008-5472.CAN-19-1077

Figure Lengend Snippet: The expression of miR-26a in RPMI8226 (A), MM.1S (B), and H929 (C) cells stably transduced with V-miR-26a-GFP or V-GFP was determined by qRT-PCR; RNU44 was the internal control. The effect of miR-26a on MM cell proliferation and migration was investigated. Expression of PARP1, c-PARP1, caspase3 and c-caspase3 in RPMI8226 and MM.1S cells transduced with V-miR-26a-GFP or V-GFP were determined by western blot. D. Predicted miR-26a target sequence on the 3’UTR of CD38. To interrupt the binding between the 3’UTR of CD38 and seed sequence of miR-26a, 4 nucleotides on the predicted region of the CD38 3’UTR were changed to the complementary sequence. A luciferase reporter assay was performed in HEK293T cells transduced with V-miR-26a-GFP or V-GFP. Expression of CD38 in RPMI8226, MM.1S, and H929 cells infected with V-miR-26a-GFP or V-GFP as determined by western blot. (*p<0.05, **p<0.01, ***p<0.001)

Article Snippet: Formalin-fixed paraffin-embedded sections were deparaffinized and then incubated with rabbit anti-CD38 polyclonal antibody (Cell Signaling Technology, #14637S) or rabbit anti-Ki-67 polyclonal antibody (Cell Signaling Technology, #9027S) or rabbit anti-cleaved caspase-3 monoblonal antibody (Cell Signaling Technology, #9664S) at 4°C overnight.

Techniques: Expressing, Stable Transfection, Transduction, Quantitative RT-PCR, Control, Migration, Western Blot, Sequencing, Binding Assay, Luciferase, Reporter Assay, Infection

Journal: Cancer research

Article Title: Targeting of CD38 by the tumor suppressor miR-26a serves as a novel potential therapeutic agent in multiple myeloma

doi: 10.1158/0008-5472.CAN-19-1077

Figure Lengend Snippet: A. CD38 siRNA and control siRNA were transfected into RPMI8226, MM.1S, and H929 cells. The expression of CD38 was verified by Western blot, the population of apoptotic cells was determined by flow cytometry, and cell viability was measured by using CellTiter-GloLuminescent Cell Viability Assay Kit. B. MM.1S cells treated with miR-26a mimic were infected with V-CD38-GFP and V-GFP; cell apoptosis and viability were determined. (*p<0.05, **p<0.01)

Article Snippet: Formalin-fixed paraffin-embedded sections were deparaffinized and then incubated with rabbit anti-CD38 polyclonal antibody (Cell Signaling Technology, #14637S) or rabbit anti-Ki-67 polyclonal antibody (Cell Signaling Technology, #9027S) or rabbit anti-cleaved caspase-3 monoblonal antibody (Cell Signaling Technology, #9664S) at 4°C overnight.

Techniques: Control, Transfection, Expressing, Western Blot, Flow Cytometry, Viability Assay, Infection

Journal: Cancer research

Article Title: Targeting of CD38 by the tumor suppressor miR-26a serves as a novel potential therapeutic agent in multiple myeloma

doi: 10.1158/0008-5472.CAN-19-1077

Figure Lengend Snippet: RPMI8226 (A) and MM.1S cells (B) transduced with V-miR-26a-GFP or V-GFP were injected subcutaneously in shoulders of SCID mice (5 mice/group). Xenograft growth was monitored for 4 weeks. Tumor diameter was measured once a week, and tumor volume was calculated. Mice were sacrificed on day 28 post-injection; xenografts were isolated and weighed. Expression of miR-26a in xenografts was determined by qRT-PCR; RNU44 was used as an internal control. Expression of Ki-67 and c-caspase3 was detected by immunohistochemistry. Protein levels of CD38 in RPMI8226 and MM.1S xenografts were measured by western blot; β-actin was used as control. (*p<0.05, **p<0.01, ***p<0.001)

Article Snippet: Formalin-fixed paraffin-embedded sections were deparaffinized and then incubated with rabbit anti-CD38 polyclonal antibody (Cell Signaling Technology, #14637S) or rabbit anti-Ki-67 polyclonal antibody (Cell Signaling Technology, #9027S) or rabbit anti-cleaved caspase-3 monoblonal antibody (Cell Signaling Technology, #9664S) at 4°C overnight.

Techniques: Transduction, Injection, Isolation, Expressing, Quantitative RT-PCR, Control, Immunohistochemistry, Western Blot

Journal: Cancer research

Article Title: Targeting of CD38 by the tumor suppressor miR-26a serves as a novel potential therapeutic agent in multiple myeloma

doi: 10.1158/0008-5472.CAN-19-1077

Figure Lengend Snippet: (A) Human naïve B cells were isolated from mormal donor peripheral blood, and the purity was deteremined by flowcytometry. (B) The naïve B cells were labeled by CFSE, cultured in 24-well plates and stimulated for 3 days with 2.6 μg/ml F(ab’)2 fragment goat anti-human IgA+IgG+IgM (H+L), 100 ng/ml recombinant human soluble CD40L, 1.0 mg/ml CpG oligodeoxynucleotide 2006, and 50 U/ml recombinant IL-2. Day 3 activated B cells were washed and cultured up to 4 days with 50 U/ml IL-2, 50ng/ml IL-6, 50 ng/ml IL-10, and 2 ng/ml IL-12 and then followed by a 3 days culture with 50ng/ml IL-6, 10ng/ml IL-15 and 500U/ml IFN-α. Cell surface CD38 and CD138 expression was determined by flowcytometry. (C) qRT-PCR was used to determine the expression of miR-26a. (D) The CD38 signal in human tonsil tissue samples was detected by immunohistochemistry, and the miR-26a signal was examined by in situ hybridization. Area 1: Germinal center B cells. Area 2: plasma cells. Scale bar = 100 μM. (E) BLIMP1 mRNA level in naïve B cells and day 10 differentiated B cells was examined by qRT-PCR. Naïve B cells were activated for 3 days and transfected with one of the BLIMP1 siRNAs, with seqeunce scrambled siRNA as control. The cells were differentiated for another 7 days and the cell surface CD38 expression was determined by flowcytometry (F) BLIMP1 expression was determined by qRT-PCR and immunoblotting (G) and the miR-26a level was determined by qRT-PCR (H). Naïve B cells were activated for 3 days and then transfected with miR-control or miR-26a mimic. The cells were cultured with IL-2, IL-6, IL-12 and IL-10, allowed to differnentiate for 4 days and then followed by a 3 days culture with IL-6, IL-15 and IFN-α. Cell surface CD38 and CD138 expression was determined by flowcytometry (I). (J) Bone marrow biopsies were obtained from the same MM patient before daratumumab treatment and after became daratumumab resistant. The CD38 expression was examined by IHC and miR-26a level was examined by ISH. Scale bar = 100μM.

Article Snippet: Formalin-fixed paraffin-embedded sections were deparaffinized and then incubated with rabbit anti-CD38 polyclonal antibody (Cell Signaling Technology, #14637S) or rabbit anti-Ki-67 polyclonal antibody (Cell Signaling Technology, #9027S) or rabbit anti-cleaved caspase-3 monoblonal antibody (Cell Signaling Technology, #9664S) at 4°C overnight.

Techniques: Isolation, Labeling, Cell Culture, Recombinant, Expressing, Quantitative RT-PCR, Immunohistochemistry, In Situ Hybridization, Clinical Proteomics, Transfection, Control, Western Blot

Journal: Cancer research

Article Title: Targeting of CD38 by the tumor suppressor miR-26a serves as a novel potential therapeutic agent in multiple myeloma

doi: 10.1158/0008-5472.CAN-19-1077

Figure Lengend Snippet: The expression of miR-26a in RPMI8226 (A), MM.1S (B), and H929 (C) cells stably transduced with V-miR-26a-GFP or V-GFP was determined by qRT-PCR; RNU44 was the internal control. The effect of miR-26a on MM cell proliferation and migration was investigated. Expression of PARP1, c-PARP1, caspase3 and c-caspase3 in RPMI8226 and MM.1S cells transduced with V-miR-26a-GFP or V-GFP were determined by western blot. D. Predicted miR-26a target sequence on the 3’UTR of CD38. To interrupt the binding between the 3’UTR of CD38 and seed sequence of miR-26a, 4 nucleotides on the predicted region of the CD38 3’UTR were changed to the complementary sequence. A luciferase reporter assay was performed in HEK293T cells transduced with V-miR-26a-GFP or V-GFP. Expression of CD38 in RPMI8226, MM.1S, and H929 cells infected with V-miR-26a-GFP or V-GFP as determined by western blot. (*p<0.05, **p<0.01, ***p<0.001)

Article Snippet: Formalin-fixed paraffin-embedded sections were deparaffinized and then incubated with rabbit anti-CD38 polyclonal antibody (Cell Signaling Technology, #14637S) or rabbit anti-Ki-67 polyclonal antibody (Cell Signaling Technology, #9027S) or rabbit anti-cleaved caspase-3 monoblonal antibody (Cell Signaling Technology, #9664S) at 4℃ overnight.

Techniques: Expressing, Stable Transfection, Transduction, Quantitative RT-PCR, Migration, Western Blot, Sequencing, Binding Assay, Luciferase, Reporter Assay, Infection

Journal: Cancer research

Article Title: Targeting of CD38 by the tumor suppressor miR-26a serves as a novel potential therapeutic agent in multiple myeloma

doi: 10.1158/0008-5472.CAN-19-1077

Figure Lengend Snippet: A. CD38 siRNA and control siRNA were transfected into RPMI8226, MM.1S, and H929 cells. The expression of CD38 was verified by Western blot, the population of apoptotic cells was determined by flow cytometry, and cell viability was measured by using CellTiter-GloLuminescent Cell Viability Assay Kit. B. MM.1S cells treated with miR-26a mimic were infected with V-CD38-GFP and V-GFP; cell apoptosis and viability were determined. (*p<0.05, **p<0.01)

Article Snippet: Formalin-fixed paraffin-embedded sections were deparaffinized and then incubated with rabbit anti-CD38 polyclonal antibody (Cell Signaling Technology, #14637S) or rabbit anti-Ki-67 polyclonal antibody (Cell Signaling Technology, #9027S) or rabbit anti-cleaved caspase-3 monoblonal antibody (Cell Signaling Technology, #9664S) at 4℃ overnight.

Techniques: Transfection, Expressing, Western Blot, Flow Cytometry, Viability Assay, Infection

Journal: Cancer research

Article Title: Targeting of CD38 by the tumor suppressor miR-26a serves as a novel potential therapeutic agent in multiple myeloma

doi: 10.1158/0008-5472.CAN-19-1077

Figure Lengend Snippet: RPMI8226 (A) and MM.1S cells (B) transduced with V-miR-26a-GFP or V-GFP were injected subcutaneously in shoulders of SCID mice (5 mice/group). Xenograft growth was monitored for 4 weeks. Tumor diameter was measured once a week, and tumor volume was calculated. Mice were sacrificed on day 28 post-injection; xenografts were isolated and weighed. Expression of miR-26a in xenografts was determined by qRT-PCR; RNU44 was used as an internal control. Expression of Ki-67 and c-caspase3 was detected by immunohistochemistry. Protein levels of CD38 in RPMI8226 and MM.1S xenografts were measured by western blot; β-actin was used as control. (*p<0.05, **p<0.01, ***p<0.001)

Article Snippet: Formalin-fixed paraffin-embedded sections were deparaffinized and then incubated with rabbit anti-CD38 polyclonal antibody (Cell Signaling Technology, #14637S) or rabbit anti-Ki-67 polyclonal antibody (Cell Signaling Technology, #9027S) or rabbit anti-cleaved caspase-3 monoblonal antibody (Cell Signaling Technology, #9664S) at 4℃ overnight.

Techniques: Transduction, Injection, Isolation, Expressing, Quantitative RT-PCR, Immunohistochemistry, Western Blot

Journal: Cancer research

Article Title: Targeting of CD38 by the tumor suppressor miR-26a serves as a novel potential therapeutic agent in multiple myeloma

doi: 10.1158/0008-5472.CAN-19-1077

Figure Lengend Snippet: (A) Human naïve B cells were isolated from mormal donor peripheral blood, and the purity was deteremined by flowcytometry. (B) The naïve B cells were labeled by CFSE, cultured in 24-well plates and stimulated for 3 days with 2.6 μg/ml F(ab’)2 fragment goat anti-human IgA+IgG+IgM (H+L), 100 ng/ml recombinant human soluble CD40L, 1.0 mg/ml CpG oligodeoxynucleotide 2006, and 50 U/ml recombinant IL-2. Day 3 activated B cells were washed and cultured up to 4 days with 50 U/ml IL-2, 50ng/ml IL-6, 50 ng/ml IL-10, and 2 ng/ml IL-12 and then followed by a 3 days culture with 50ng/ml IL-6, 10ng/ml IL-15 and 500U/ml IFN-α. Cell surface CD38 and CD138 expression was determined by flowcytometry. (C) qRT-PCR was used to determine the expression of miR-26a. (D) The CD38 signal in human tonsil tissue samples was detected by immunohistochemistry, and the miR-26a signal was examined by in situ hybridization. Area 1: Germinal center B cells. Area 2: plasma cells. Scale bar = 100 μM. (E) BLIMP1 mRNA level in naïve B cells and day 10 differentiated B cells was examined by qRT-PCR. Naïve B cells were activated for 3 days and transfected with one of the BLIMP1 siRNAs, with seqeunce scrambled siRNA as control. The cells were differentiated for another 7 days and the cell surface CD38 expression was determined by flowcytometry (F) BLIMP1 expression was determined by qRT-PCR and immunoblotting (G) and the miR-26a level was determined by qRT-PCR (H). Naïve B cells were activated for 3 days and then transfected with miR-control or miR-26a mimic. The cells were cultured with IL-2, IL-6, IL-12 and IL-10, allowed to differnentiate for 4 days and then followed by a 3 days culture with IL-6, IL-15 and IFN-α. Cell surface CD38 and CD138 expression was determined by flowcytometry (I). (J) Bone marrow biopsies were obtained from the same MM patient before daratumumab treatment and after became daratumumab resistant. The CD38 expression was examined by IHC and miR-26a level was examined by ISH. Scale bar = 100μM.

Article Snippet: Formalin-fixed paraffin-embedded sections were deparaffinized and then incubated with rabbit anti-CD38 polyclonal antibody (Cell Signaling Technology, #14637S) or rabbit anti-Ki-67 polyclonal antibody (Cell Signaling Technology, #9027S) or rabbit anti-cleaved caspase-3 monoblonal antibody (Cell Signaling Technology, #9664S) at 4℃ overnight.

Techniques: Isolation, Labeling, Cell Culture, Recombinant, Expressing, Quantitative RT-PCR, Immunohistochemistry, In Situ Hybridization, Transfection, Western Blot