syntaxin 5  (Santa Cruz Biotechnology)

Journal: Traffic (Copenhagen, Denmark)

Article Title: Acute GARP Depletion Disrupts Vesicle Transport, Leading to Severe Defects in Sorting, Secretion and O ‐Glycosylation

doi: 10.1111/tra.70003

Figure Lengend Snippet: List of primary and secondary antibodies.

Article Snippet: STX5 , Santa Cruz, #sc‐365124 , Mouse , — , 1:100.

Techniques: Transduction

Journal: Traffic (Copenhagen, Denmark)

Article Title: Acute GARP Depletion Disrupts Vesicle Transport, Leading to Severe Defects in Sorting, Secretion and O ‐Glycosylation

doi: 10.1111/tra.70003

Figure Lengend Snippet: v‐SNARE GS15 is mislocalized in VPS54‐depleted cells. (A) RPE1 VPS54‐mAID cells were treated with AA as indicated, and cell lysates were probed with (top panel) anti‐GS15 (A) and anti‐GS28 (D). β‐Actin was used as a loading control. The bottom panels on (A) and (D) are the quantification of the blots from three independent experiments. (B) Airyscan microscopy of RPE1 VPS54‐mAID cells untreated or treated with AA for 3 h and co‐stained for GS15 and P230. (C) Colocalization analysis of GS15 and P230 of ≥ 30 cells was done by calculating Pearson's correlation coefficient. Statistical significance was calculated using a paired t ‐test. ** p ≤ 0.01. (F) WB analysis of RPE1 VPS54‐mAID cells treated with AA and probed with antibodies to STX5, STX6, STX10, VAMP4 and VTI1A, respectively. β‐Actin was used as a loading control.

Article Snippet: STX5 , Santa Cruz, #sc‐365124 , Mouse , — , 1:100.

Techniques: Control, Microscopy, Staining

Journal: Traffic (Copenhagen, Denmark)

Article Title: Acute GARP Depletion Disrupts Vesicle Transport, Leading to Severe Defects in Sorting, Secretion and O ‐Glycosylation

doi: 10.1111/tra.70003

Figure Lengend Snippet: Rapid VPS54 depletion results in the accumulation of GARP‐dependent vesicles and alteration of TGN morphology. (A) Transmission Electron Microscopy of high‐pressure frozen RPE1 VPS54‐mAID cells grown on sapphire discs before and after 3 h of AA treatment. “G” indicates Golgi stacks. Arrowheads point to vesicle‐like structures. Arrows indicate the enlarged vacuolar structures accumulated near the Golgi. Asterisks indicate the autophagosomes. Scale bar, 500 nm. (B) The graph represents the quantification of the total number of vesicles around the Golgi before and after 3 h of AA treatment. (C) Schematic of the cellular fractionation experiment to prepare P30 (Golgi), and P100 (Vesicle) fractions from control and 3 h AA treated groups. (D) WB analysis of TGN localized proteins (TGN46, CI‐MPR and CD‐MPR) in Golgi and vesicle fractions. (E) WB analysis of Golgi enzymes (B4GALT1, MGAT1, C1GALT1, GALNT2 and CPD) in Golgi and vesicle fractions. (F) WB analysis of SNAREs (STX5, GS15, STX10, STX6) in Golgi and vesicle fractions.

Article Snippet: STX5 , Santa Cruz, #sc‐365124 , Mouse , — , 1:100.

Techniques: Transmission Assay, Electron Microscopy, Cell Fractionation, Control