Journal: Plant Direct

Article Title: A chemical genetic screen with the EXO70 inhibitor Endosidin2 uncovers potential modulators of exocytosis in Arabidopsis

doi: 10.1002/pld3.592

Figure Lengend Snippet: Endosidin2 (ES2) inhibits Arabidopsis root growth in a dose‐dependent manner. (a) Seven‐day‐old light‐grown seedlings of PIN2‐GFP and Col‐0 grown on solid ½ MS media containing various concentrations of ES2. Representative seedlings are displayed. Scale bar = 1 cm. (b) Growth curve of root lengths for seedlings after ES2 treatment. (c) Box plot of root lengths for 7‐day‐old seedlings with ES2 dose series treatment, with whiskers displaying the minimum and maximum root length in each group. Statistical analysis was performed with one‐way ANOVA, with Dunnett multiple comparison with the control group. ***denotes p < 0.001, **** denotes p < 0.0001, ns denotes not significant. N = 45–60 seedlings for PIN2‐GFP , and 58–74 seedlings for Col‐0 , from 3 biological replicates.

Article Snippet: Endosidin2 [(E)‐3‐Fluoro‐N′‐(4‐hydroxy‐3‐iodo‐5‐methoxybenzylidene)‐benzohydrazide] (Molecular weight 414.17 Da) was synthesized from the reaction between3‐fluorophenylhydrazide and 5‐iodovanillin at the Department of Chemistry, Purdue University.

Techniques: Comparison, Control

Journal: Cancers

Article Title: Exo70 Promotes the Invasion of Pancreatic Cancer Cells via the Regulation of Exosomes

doi: 10.3390/cancers16020336

Figure Lengend Snippet: Knockdown of Exo70 caused intracellular aggregation of MVBs. ( A , B ) Transmission electron microscopy images of MVEs in PC cells (A818-4 and PANC-1) stably expressing control or Exo70 knockdown or in cells treated with DMSO or ES2. Scale bars = 5 μm. Black arrows indicate MVEs. Dashed rectangles indicate regions zoomed in the respective lower panels. ( C , D ) Quantification of the number of MVEs and the ILV numbers of each MVE in PC cells (A818-4 and PANC-1). ( E , F ) Immunofluorescence staining of CD63 in PC cells (A818-4 and PANC-1) stably expressing the control or Exo70knockdown. Scale bars = 5 μm. * p < 0.05, ** p < 0.01, **** p < 0.0001.

Article Snippet: The following antibodies and reagents were used for this study: anti-β-actin (Sigma-Aldrich, St. Louis, MO, USA, cat#a3854), anti-ALIX (Abcam, Cambridge, UK, cat#ab186429), anti-CD63 (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA, cat#sc-5275), anti-CD81 (Proteintech, Wuhan, China, cat#66866-1-lg), anti-CD9 (Proteintech, cat#60232-1-lg), anti-Tsg101 (Abcam, cat#ab133586), anti-Exo70 (Abcam, cat#ab118792), anti-F-actin (Cytoskeleton, Inc., Denver, CO, USA; cat#PHDR1), anti-PD-L1 (Cell Signaling, Danvers, MA, USA, cat# 13684), Endosidin2 (ES2) (Cayman Chemical, Ann Arbor, MI, USA, cat#21888) and PKH67 (Sigma-Aldrich, cat#MINI67).

Techniques: Knockdown, Transmission Assay, Electron Microscopy, Stable Transfection, Expressing, Control, Immunofluorescence, Staining

Journal: Cancers

Article Title: Exo70 Promotes the Invasion of Pancreatic Cancer Cells via the Regulation of Exosomes

doi: 10.3390/cancers16020336

Figure Lengend Snippet: Exo70 promoted tumor metastasis in mice. ( A , E ) Flow diagram of the mouse experiments. ( B , F ) Mouse weights. ( C ) Bioluminescent imaging for A818-4-luc orthotopic xenograft colon tumors in the control and ES2 group (n = 5 per group). ( D ) Comparison of the liver bioluminescence between the control and ES2 group. ( G ) Bioluminescent imaging for A818-4-luc tumors in the sh-Ctrl, sh-Exo70 and sh-Exo70 + Exo groups (n = 5 per group). ( H ) Images of mouse livers and metastatic tumors. ( I ) Comparison of the bioluminescence between the sh-Ctrl, sh-Exo70 and sh-Exo70 + Exo groups. ( J ) Tumor number statistics. ns p > 0.05, * p < 0.05, ** p < 0.01.

Article Snippet: The following antibodies and reagents were used for this study: anti-β-actin (Sigma-Aldrich, St. Louis, MO, USA, cat#a3854), anti-ALIX (Abcam, Cambridge, UK, cat#ab186429), anti-CD63 (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA, cat#sc-5275), anti-CD81 (Proteintech, Wuhan, China, cat#66866-1-lg), anti-CD9 (Proteintech, cat#60232-1-lg), anti-Tsg101 (Abcam, cat#ab133586), anti-Exo70 (Abcam, cat#ab118792), anti-F-actin (Cytoskeleton, Inc., Denver, CO, USA; cat#PHDR1), anti-PD-L1 (Cell Signaling, Danvers, MA, USA, cat# 13684), Endosidin2 (ES2) (Cayman Chemical, Ann Arbor, MI, USA, cat#21888) and PKH67 (Sigma-Aldrich, cat#MINI67).

Techniques: Imaging, Control, Comparison