Journal: Cell Reports

Article Title: Mechanistic insights into COVID-19 by global analysis of the SARS-CoV-2 3CL pro substrate degradome

doi: 10.1016/j.celrep.2021.109892

Figure Lengend Snippet:

Article Snippet: The primary antibodies and dilutions used were: mouse monoclonal anti-SARS-CoV-2 nucleocapsid antibody (1:1,000, Invitrogen, MA5-29981, RRID: AB_2785780 ); rabbit anti-SARS-CoV-1 3CL pro antibody (1:2000, Rockland, 200-401-A51, RRID: AB_828457 ); rabbit polyclonal anti-RPAP1 antibody (1:1,000, Proteintech, 15138-1-AP, RRID: AB_2301137 ); mouse monoclonal anti-PTBP1 antibody (1:500, Biolegend, 630101, 3H7, RRID: AB_2171285 ); rabbit polyclonal anti-MAP4K5 antibody (1:1,000, Cusabio, CSB-PA013440DSR2HU, RRID: AB_2892084 ); rabbit polyclonal anti-CREB1 antibody (1:1,000, Abclonal, A11989, RRID: AB_2758916 ); rabbit polyclonal anti-YAP1 antibody (1:1,000, Abclonal, A11430, RRID: AB_2758556 ); rabbit polyclonal anti-FYCO1 antibody (1:1,000, Cusabio, CSB-PA866262LA01HU, RRID: AB_2892085 ); rabbit polyclonal anti-FAF1 antibody (1:1,000, Abclonal, A2921, RRID: AB_2764739 ); goat polyclonal anti-Gal8 antibody (1:400, R&D Systems, AF1305, RRID: AB_2137229 ); rabbit polyclonal anti-KPNA3 (IMA4) antibody (1:1,000, Abclonal, A8347, RRID: AB_2770124 ); rabbit polyclonal anti-NUP107 antibody (1:1,000, Abclonal, A13110, RRID: AB_2759959 ); mouse monoclonal anti-IRS2 antibody (1:300, R&D Systems, MAB6347, 676415, RRID: AB_10992928 ); mouse monoclonal anti-FLAG M2 antibody (1:10,000, Sigma, F3165, RRID: AB_259529 ); mouse monoclonal anti-β-tubulin antibody (1:2000, AbLab, 21-0018-00, clone BT7R); mouse monoclonal anti-β-actin antibody (1:1,000, Abcam, ab8226, RRID: AB_306371 ); rabbit monoclonal anti-β-actin antibody (1:200, Abcam, ab115777, RRID: AB_10899528 ).

Techniques: Virus, Recombinant, Sequencing, Fluorescence, Modification, Protease Inhibitor, Polymer, Staining, Blocking Assay, Binding Assay, Activity Assay, Western Blot, Synthesized, Software, Control, Targeted Proteomics, Microscopy, Spectrophotometry, Mass Spectrometry

Journal: Cancer Medicine

Article Title: Gene expression of the IGF pathway family distinguishes subsets of gastrointestinal stromal tumors wild type for KIT and PDGFRA

doi: 10.1002/cam4.57

Figure Lengend Snippet: Target gene expression relative to GAPDH in GIST subtypes

Article Snippet: Additional targets were detected using commercial TaqMan Gene Expression assays (Applied Biosystems by Life Technologies): IGF1R (64-bp amplicon) assay Hs00609566_m1, IGF1 (63 bp) Hs01547657_m1, IGF2 (81 bp) Hs01005963_m1, INSR (76 bp) Hs00965956_m1, IRS1 (insulin receptor substrate [IRS]) (69 bp) Hs00178563_m1, IRS2 (70 bp) Hs00275843_s1, CDH2 [Cadherin (CDH)] (78 bp) Hs00983062_m1, neurofilament, light polypeptide (NEFL) (71 bp) Hs00196245_m1, LHX2 (LIM homeobox [LHX]) (64 bp) Hs01104717_m1, KIRREL3 kin of IRRE like (KIRREL) (68 bp) Hs01123170_m1, CD34 (61 bp) Hs00990730_m1, and HIF1A (62 bp) Hs00936371_m1.

Techniques: Targeted Gene Expression

Journal: Ecotoxicology and environmental safety

Article Title: Azadirachtin exposure inhibit ovary development of Spodoptera litura (Lepidoptera: Noctuidae) by altering lipids metabolism event and inhibiting insulin signaling pathways.

doi: 10.1016/j.ecoenv.2023.115151

Figure Lengend Snippet: Fig. 6. AZA exposure led to insulin resistance- like phenotypes in ovary of S.litura. (A) INS level in ovary. (B) Glu level in ovary. (C) Gly level in ovary. (D) FFA level in ovary. (E) Western blot for insulin signal-related proteins (PI3K, IRS1, P-IRS1 (ser307), IRS2, P-IRS2 (ser731), AKT and P-AKT). (F) The bar graph showed the relative protein abundances fol lowed western blot by normalizing the signals to β-Actin via Image J software. Data was rep resented as mean ± SEM (n = 3), significance was determined by Student’s unpaired t tests. Values significantly differ at * ** p-value < 0.001.

Article Snippet: The membranes were blocked with 5 % skim milk in TBST buffer (Tris-HCl, pH 7.5, 150 mM NaCl, and 0.1 % Tween 20) for 2 h. After washing three times with PBST, the membranes were incubated with primary antibodies against the following proteins, IRS1 (PB0221, Boster, Wuhan, China), P-IRS1 (ser307) (abs130899, Absin, Shanghai, China), IRS2 (PB0222, Boster, Wuhan, China), P-IRS2 (ser731) (bs-10998R, Bioss, Beijing, China), AKT (bs-6951R, Bioss, Beijing, China), P-AKT (ser505) (#4054, Cell Signaling Technology, USA), PI3K p85 (#4257, Cell Signaling Technology, USA) and the reference proteins (β-Actin) (AF003, Beyotime, Shanghai, China), respectively.

Techniques: Western Blot, Software

Journal: PLoS ONE

Article Title: Recurrent Amplification at 13q34 Targets at CUL4A , IRS2 , and TFDP1 As an Independent Adverse Prognosticator in Intrahepatic Cholangiocarcinoma

doi: 10.1371/journal.pone.0145388

Figure Lengend Snippet: Description of western blot and immunohistochemical antibodies.

Article Snippet: The IRS2 overexpression vector (pCMV Entry-IRS2-Myc-DDK, Cat. No. RC212413) and specific siRNAs targeting IRS2 , CUL4A and TFDP1 were purchased from Origene (Origene, MD, USA).

Techniques: Western Blot, Immunohistochemical staining

Journal: PLoS ONE

Article Title: Recurrent Amplification at 13q34 Targets at CUL4A , IRS2 , and TFDP1 As an Independent Adverse Prognosticator in Intrahepatic Cholangiocarcinoma

doi: 10.1371/journal.pone.0145388

Figure Lengend Snippet: (a) SNU1079 cells were transiently transfected with expression vector (SNU1079-IRS2). After 48 hours, expression of IRS2 was determined by immunofluorescence detection of IRS2 protein (left panel, green) and DAPI (middle panel, blue). MERGE contains the combined image of both GFP and DAPI staining (right panel). Percentage of IRS2-positive SNU1079 cells was normalized to DAPI stained nuclei. Images were obtained at a magnification of × 100. (b) Total cell lysates were analyzed for IRS2 protein levels by Western blotting. Anti-DDK tag antibody was used to reveal the extent of IRS2 overexpression. (c) The effect of IRS2 overexpression on growth potential was determined by Western blotting using anti-PCNA antibody and manual counting of total cell numbers. (d) SNU1079-IRS2 cells were subjected to invasion and migration assays, and showed a greater degree of migration and invasion, compared with control cells. Graphs represent data from one of three independent experiments with similar results. Data represent means ± standard deviations for three independent experiments.

Article Snippet: The IRS2 overexpression vector (pCMV Entry-IRS2-Myc-DDK, Cat. No. RC212413) and specific siRNAs targeting IRS2 , CUL4A and TFDP1 were purchased from Origene (Origene, MD, USA).

Techniques: Transfection, Expressing, Plasmid Preparation, Immunofluorescence, Staining, Western Blot, Over Expression, Migration

Journal: PLoS ONE

Article Title: Recurrent Amplification at 13q34 Targets at CUL4A , IRS2 , and TFDP1 As an Independent Adverse Prognosticator in Intrahepatic Cholangiocarcinoma

doi: 10.1371/journal.pone.0145388

Figure Lengend Snippet: Copy number changes of target genes at 13q34.

Article Snippet: The IRS2 overexpression vector (pCMV Entry-IRS2-Myc-DDK, Cat. No. RC212413) and specific siRNAs targeting IRS2 , CUL4A and TFDP1 were purchased from Origene (Origene, MD, USA).

Techniques:

Journal: PLoS ONE

Article Title: Recurrent Amplification at 13q34 Targets at CUL4A , IRS2 , and TFDP1 As an Independent Adverse Prognosticator in Intrahepatic Cholangiocarcinoma

doi: 10.1371/journal.pone.0145388

Figure Lengend Snippet: Independent predictive factors of disease-free survival by multivariate analysis.

Article Snippet: The IRS2 overexpression vector (pCMV Entry-IRS2-Myc-DDK, Cat. No. RC212413) and specific siRNAs targeting IRS2 , CUL4A and TFDP1 were purchased from Origene (Origene, MD, USA).

Techniques:

Journal: PLoS ONE

Article Title: Recurrent Amplification at 13q34 Targets at CUL4A , IRS2 , and TFDP1 As an Independent Adverse Prognosticator in Intrahepatic Cholangiocarcinoma

doi: 10.1371/journal.pone.0145388

Figure Lengend Snippet: Positive nuclear staining was observed for CUL 4A and TFDP1, and cytoplasmic staining for IRS2 in tumor cells with amplification of target genes (a, e, i), compared with normal bile ductules in the adjacent portal tracts (b, f, j). Loss of expression for CUL 4A, IRS2, and TFDP1 (c, g, k) was observed in tumor cells containing a deletion of target genes, in contrast to their normal counterparts (d, h, l). Original magnification: × 200.

Article Snippet: The IRS2 overexpression vector (pCMV Entry-IRS2-Myc-DDK, Cat. No. RC212413) and specific siRNAs targeting IRS2 , CUL4A and TFDP1 were purchased from Origene (Origene, MD, USA).

Techniques: Staining, Amplification, Expressing