Journal: Environmental Health Perspectives

Article Title: Effects of Early Life Oral Arsenic Exposure on Intestinal Tract Development and Lipid Homeostasis in Neonatal Mice: Implications for NAFLD Development

doi: 10.1289/EHP12381

Figure Lengend Snippet: The morphological, cellular, and molecular changes of the small intestine in hUGT1 neonatal mice after a single-dose iAs treatment at 10 mg / kg BW. Neonatal hUGT1 mice (12–14 d old) were orally treated with control vehicle (C) or iAs ( 10 mg / kg BW) for various time points, and the mice were sacrificed at 14 d of age. (A) Mouse BW at 48 h after treatment ( n = 17 , 24, mean ± SEM , two-tailed Student’s t -test). (B–D) Gross photo of small intestines at 48 h posttreatment, small intestine length and weight ( n = 4 , 5, mean ± SEM , two-tailed Student’s t -test). (E) Gut transit time at 24 h after iAs treatment ( n = 5 , 6, mean ± SEM , two-tailed Student’s t -test). (F) Immunostaining of Ki67. Ki 67 + cells were enumerated and described as Ki 67 + cells per crypt ( n = 3 , mean ± SEM , one-way ANOVA). Scale bars: 50 μ m (upper) and 20 μ m (lower). (G) Western blot analysis of intestinal LGR5, PCNA, and SIS at 24 and 48 h after iAs treatment. (H) RT-qPCR ( n = 3 , mean ± SEM ). * p < 0.05 , ** p < 0.01 , *** p < 0.001 . Corresponding numeric data and individual p -values are listed in Excel Table S1. Note: Akp3, alkaline phosphatase 3; ANOVA, analysis of variance; BW, body weight; Cph, cyclophilin; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; iAs, inorganic arsenic; Ki 67 + , Ki67-positive; LGR5, leucine-rich repeat-containing G protein-coupled receptor 5; PCNA, proliferating cell nuclear antigen; RT-qPCR, real-time quantitative polymerase chain reaction; SEM, standard error of the mean; SI, small intestine; SIS, sucrase isomaltase.

Article Snippet: The slides were then incubated with anti-Ki67 antibody (Abcam; Catalog no. Ab-149; dilution factor, 1:100) overnight at 4°C.

Techniques: Two Tailed Test, Immunostaining, Western Blot, Quantitative RT-PCR, Real-time Polymerase Chain Reaction

Journal: Environmental Health Perspectives

Article Title: Effects of Early Life Oral Arsenic Exposure on Intestinal Tract Development and Lipid Homeostasis in Neonatal Mice: Implications for NAFLD Development

doi: 10.1289/EHP12381

Figure Lengend Snippet: The lipid levels in enterocytes isolated from iAs-treated hUGT1 neonatal mice, as well as the morphological, cellular, and molecular changes of hUGT1 neonatal mice following treatment with OA. Neonatal hUGT1 mice were orally treated with control vehicle (C) or iAs at 10 mg / kg BW. Small intestines were collected at 24 h and 48 h after treatment. Images were examined using a 20 × Plan-Apochromat objective (numeric aperture, 0.8) on an upright Imager A2 microscope (Zeiss) with an Axiocam 506 color camera and ZEN2012 imaging software. Scale bars: 50 μ m (upper) and 20 μ m (lower). (A) Representative micrograph of H&E staining ( n = 3 ). (B) Representative micrograph of Oil red O staining ( n = 3 ). (C) Photos of aliquots of enterocyte lysates. (D–F) TGs ( n = 7 , 4, mean ± SEM , two-tailed Student’s t -test), FFAs ( n = 6 , mean ± SEM , two-tailed Student’s t -test), and cholesterol ( n = 3 , mean ± SEM ) content in enterocytes. (G) Schematic description of the impact of iAs on lipids and IEC cell proliferation and maturation. (H–K) Neonatal hUGT1 mice were orally treated with control vehicle (C) or OA at 10 g / kg BW. Small intestines were collected at 24 h and 48 h. (H) Small intestine weight at 48 h after treatment ( n = 8 , 5, mean ± SEM , two-tailed Student’s t -test). (I) RT-qPCR at 48 h posttreatment, examining Sis and Akp3 gene expression ( n = 8 , 5, mean ± SEM , two-tailed Student’s t -test). (J) Western blot analysis of LGR5 and SIS. (K) Immunostaining of Ki67, Ki 67 + cells were enumerated and described as Ki 67 + cells per crypt ( n = 3 , mean ± SEM , one-way ANOVA). * p < 0.05 , ** p < 0.01 , *** p < 0.001 , **** p < 0.0001 . Corresponding numeric data and individual p -values are listed in Excel Table S2. Note: Akp3, alkaline phosphatase 3; ANOVA, analysis of variance; BMK, breast milk; BW, body weight; Cph, cyclophilin; FFAs, free fatty acids; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; H&E, hematoxylin and eosin; iAs, inorganic arsenic; IEC, intestinal epithelial cell; Ki 67 + , Ki67-positive; LGR5, leucine-rich repeat-containing G protein-coupled receptor 5; OA, oleic acid; RT-qPCR, real-time quantitative polymerase chain reaction; SEM, standard error of the mean; SI, small intestine; SIS, sucrase isomaltase; TGs, triglycerides.

Article Snippet: The slides were then incubated with anti-Ki67 antibody (Abcam; Catalog no. Ab-149; dilution factor, 1:100) overnight at 4°C.

Techniques: Isolation, Microscopy, Imaging, Software, Staining, Two Tailed Test, Quantitative RT-PCR, Expressing, Western Blot, Immunostaining, Real-time Polymerase Chain Reaction

Journal: eLife

Article Title: An engineered transcriptional reporter of protein localization identifies regulators of mitochondrial and ER membrane protein trafficking in high-throughput CRISPRi screens

doi: 10.7554/eLife.69142

Figure Lengend Snippet:

Article Snippet: Primary antibodies used in imaging include the following: anti-V5 (Mouse, Invitrogen R960); anti-TOMM20 (Rabbit, Abcam ab186735); anti-GRASP65 (Mouse, Santa Cruz sc-374423); anti-CANX (Rabbit, Thermo Fisher PA5-34754); anti-PEX14 (Rabbit, Proteintech 10594-1-AP); anti-RCN2 (Rabbit, Thermo Fisher PA5-56542); anti-AKAP1 (Mouse, Santa Cruz sc-135824); and anti-MAVS (Mouse, Santa Cruz sc-166583).

Techniques: Immunofluorescence, Western Blot, Recombinant, Sequencing, Selection, Random Hexamer, Derivative Assay, BIA-KA, Software, Imaging, Reverse Transcription, Protease Inhibitor