Journal: Clinical and Translational Medicine

Article Title: Muscle‐specific gene editing improves molecular and phenotypic defects in a mouse model of myotonic dystrophy type 1

doi: 10.1002/ctm2.70227

Figure Lengend Snippet: Improvement of bone health and body composition parameters in MyoAAV–C3/384‐treated homozygous DMSXL mice. A group of DMSXL male and female homozygous mice at age P5 received a systemic injection of MyoAAV vectors carrying Cas9 and sgC3/384 pair ( n = 7), and underwent whole‐body DEXA analysis at 16 weeks of age. Control animals were wild‐types (WT, n = 16) and untreated DMSXL homozygous mice (NT, n = 14) of both sexes. All data are represented as mean ± SEM and individual data points; data from male (open symbols) and female (filled symbols) mice are pooled. (A–C) Bone health parameters: bone mineral density (BMD) (A), bone mineral content (BMC) (B) and bone area (C). (A) BMD: MyoAAV–C3/384 vs. NT, unpaired t ‐test with Welch's correction, t = 2.595, df = 14, * p = .02. (B) BMC: MyoAAV–C3/384 vs. NT, unpaired t ‐test with Welch's correction, t = 3.371, df = 15, ** p = .004. (C) Bone area, WT vs. NT, Mann–Whitney test, U = 53, * p = .013; MyoAAV–C3/384 vs. NT, unpaired t ‐test with Welch's correction, t = 4.25, df = 16, *** p = .0006. (D–F) Body composition parameters: fat tissue percentage (D), lean tissue percentage (E) and fat/lean mass ratio (adiposity index, F). (D) Fat tissue %: WT vs. NT, Mann–Whitney test, U = 46, ** p = .0052; MyoAAV–C3/384 vs. NT, unpaired t ‐test with Welch's correction, t = 2.684, df = 17.14, * p = .015. (E) Lean tissue %: WT vs. NT, Mann–Whitney test, U = 45, ** p = .0045; MyoAAV–C3/384 vs. NT, unpaired t ‐test with Welch's correction, t = 2.684, df = 17.14, * p = .015. (F) Adiposity index, WT vs. NT, unpaired t ‐test with Welch's correction, t = 3.265, df = 15.37, ** p = .0051; MyoAAV–C3/384 vs. NT, unpaired t ‐test with Welch's correction, t = 2.643, df = 16.41, * p = .017.

Article Snippet: Bone health and body composition were comprehensively evaluated at the age of 4 months using Dual Energy X‐ray Absorptiometry (DEXA) analyser (The PARAMETER™ XPERT® 40 Cabinet X‐ray System, T00261_A 9.2019; Kubtec Scientific, 111 Research Drive Stratford, CT 06615, USA).

Techniques: Injection, Control, MANN-WHITNEY

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: E-Selectin-Dependent Inflammation and Lipolysis in Adipose Tissue Exacerbate Steatosis-to-NASH Progression via S100A8/9

doi: 10.1016/j.jcmgh.2021.08.002

Figure Lengend Snippet: Antibodies

Article Snippet: S100A8/A9 (mouse) , 1:500 (staining) 1:5000 (WB) , Novus Biologicals.

Techniques: Staining

Journal: Drug Metabolism and Disposition

Article Title: UDP-Glucuronosyltransferase (UGT) 1A9-Overexpressing HeLa Cells Is an Appropriate Tool to Delineate the Kinetic Interplay between Breast Cancer Resistance Protein (BRCP) and UGT and to Rapidly Identify the Glucuronide Substrates of BCRP

doi: 10.1124/dmd.111.041467

Figure Lengend Snippet: TABLE 1

Article Snippet: UGT1A9 antibody was purchased from Abnova (Walnut, CA). pCMV6_XL4 vector carrying the UGT1A9 gene was from Origene (Rockville, MD). siRNA of UGT1A9 and scrambled siRNA were purchased from Ambion (Austin, TX). siRNA of MRP2 or MRP3 and 3-(6-isobutyl-9-methoxy-1,4-dioxo-1,2,3,4,6,7,12,12 a -octahydropyrazino[1′,2′:1,6]pyrido[3,4- b ]indol-3-yl)-propionic acid tert -butyl ester (Ko143) were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA).

Techniques:

Journal: Oncogene

Article Title: Osteopontin signaling upregulates cyclooxygenase-2 expression in tumor-associated macrophages leading to enhanced angiogenesis and melanoma growth via α9β1 integrin.

doi: 10.1038/onc.2013.184

Figure Lengend Snippet: Figure 3. OPN upregulates COX-2 expression and COX-2-dependent PGE2 production in an autocrine manner in macrophages via a9 integrin. (a) RAW264.7 cells were supplemented with CM of melanoma (B16F10 or A375), pre-incubated with OPN neutralizing antibody (5 mg/ml) and expression of COX-2 was analyzed by western blot (upper panel). RAW264.7 cells were supplemented with CM of melanoma (B16F10 or A375) cells transfected with OPN siRNA (60 pmol/ml). COX-2 expression was analyzed by western blot (lower panel). Actin was used as loading control. (b) RAW264.7 cells were transfected with OPN siRNA (60 pmol/ml) and then treated with CM of B16F10 and expressions of COX-2 and OPN were analyzed by western blot (upper panel). Peritoneal macrophages (pMac) isolated from Opn þ / þ (^ þ / þ) and Opn / (^ / ) mice were treated with CM of B16F10 and COX-2 expression was analyzed by western blot (lower panel). (c) RAW264.7 cells or pMac were treated with various doses of OPN and COX-2 expression was analyzed by western blot (upper and lower panels). (d) RAW264.7 cells were transfected with OPN cDNA, OPN siRNA (60 pmol/ml) or OPN siRNA-transfected cells were cotransfected with OPN cDNA or treated with human or mouse OPN. The expressions of COX-2 and OPN were analyzed by western blot (left panel). RAW264.7 cells were treated with OPN (0-1 mM). The level of PGE2 was estimated by EIA and represented in the form of bar graph. The error bar represents s.e.m., *Po0.05 versus control (right panel). The data represent three experiments exhibiting similar results. (e) RAW264.7 cells were transfected with a9 integrin siRNA (40 pmol/ml) or pretreated with RGD (10 mM) or control peptide (10 mM) and then treated with OPN (0.5 mM) and COX-2 expression was analyzed by western blot (upper panel). Expression of a9 integrin in cells transfected with its specific siRNA was analyzed (lower panel). Actin was used as loading control.

Article Snippet: Small interfering RNA RAW264.7 cells were transfected with siRNA that specifically targeting OPN (siGENOMESMARTpool mouse SPP1, Dharmacon International, Lafayette, CO, USA), CD44 (CD44 siRNA), integrin a9 (integrin a9 siRNA) and COX-2 (COX-2 siRNA) (Santa Cruz Biotechnology, Santa Cruz, CA, USA) with lipofectamine 2000.

Techniques: Expressing, Incubation, Western Blot, Transfection, Control, Isolation

Journal: Oncogene

Article Title: Osteopontin signaling upregulates cyclooxygenase-2 expression in tumor-associated macrophages leading to enhanced angiogenesis and melanoma growth via α9β1 integrin.

doi: 10.1038/onc.2013.184

Figure Lengend Snippet: Figure 7. OPN-activated macrophages induce melanoma cell migration. (A) RAW264.7 cells treated with either OPN alone or pretreated with NS-398 or Etoricoxib and then treated with OPN and used in lower chamber, whereas melanoma cells were used in the upper chamber. After 12 h, migrated cells were stained and photographed (left panel). Bar graph represents the migration of melanoma cells toward OPN-activated macrophage. The error bar represents s.e.m., *Po0.001 versus untreated (Un) macrophages, ##Po0.001 versus OPN-activated macrophages (right panel). (B, C) Infiltrating macrophages (CD68 positive) in malignant melanoma are OPN and COX-2 positive and correlates with tumor angiogenesis. Histopathological (H&E; a, b) and immunohistochemical staining of peripheral normal and malignant melanoma specimens with anti-CD68 (red; c, d), co-staining with anti-CD68, anti-OPN and DAPI (e, f; white arrow indicates OPN-positive macrophages) and anti- CD31 (red; g, h). (C) Colocalization of COX-2 (green) and macrophages (CD68, red) was observed in normal peripheral and malignant melanoma specimens (a–h). (D) Schematic representation of tumor-educated OPN-activated macrophages via a9 integrin regulates ERK- and p38-dependent AP-1 activation leading to enhanced COX-2 expression and PGE2 production, which in turn upregulates melanoma growth and angiogenesis.

Article Snippet: Small interfering RNA RAW264.7 cells were transfected with siRNA that specifically targeting OPN (siGENOMESMARTpool mouse SPP1, Dharmacon International, Lafayette, CO, USA), CD44 (CD44 siRNA), integrin a9 (integrin a9 siRNA) and COX-2 (COX-2 siRNA) (Santa Cruz Biotechnology, Santa Cruz, CA, USA) with lipofectamine 2000.

Techniques: Migration, Staining, Immunohistochemical staining, Activation Assay, Expressing