Journal: Autophagy

Article Title: Mammalian BCAS3 and C16orf70 associate with the phagophore assembly site in response to selective and non-selective autophagy

doi: 10.1080/15548627.2021.1874133

Figure Lengend Snippet: The BCAS3-C16orf70 complex binds to PtdIns3P in vitro . (A) CBB staining of purified GFP-His, GFP-WIPI1-His, GFP-BCAS3-His/GFP-C16orf70-His and GFP-BCAS3 R401A -His/GFP-C16orf70-His. (B) Schematic diagram of the liposome flotation assay. After incubating the liposome solution with purified proteins, OptiPrep was added to a final concentration of 35% and then 30% OptiPrep and buffer were sequentially layered on top. After centrifugation, the Bottom (B), Middle (M) and Top (T) fractions are collected and analyzed by western blotting. (C) Flotation assays using the purified proteins and liposomes containing the indicated phosphoinositides were performed. GFP-His and GFP-WIPI1-His were detected with an anti-GFP antibody, and GFP-BCAS3-His was detected with an anti-BCAS3 antibody. (D) Flotation assays using liposomes with various concentrations of PtdIns3P were performed. GFP-BCAS3-His was detected with an anti-BCAS3 antibody

Article Snippet: After four 10 min water washes, the proteins were detected using CBB Stain One (nakalai tesque, 04543–51).

Techniques: In Vitro, Staining, Purification, Concentration Assay, Centrifugation, Western Blot, Liposomes

Journal: Inorganic Chemistry

Article Title: Azolato-Bridged Dinuclear Platinum(II) Complexes Exhibit Androgen Receptor-Mediated Anti-Prostate Cancer Activity

doi: 10.1021/acs.inorgchem.4c01093

Figure Lengend Snippet: 5-H-Y binds directly to androgen receptor (AR). (A) Coomassie Brilliant Blue stain confirming that bovine serum albumin forms aggregates in the presence of 5-H-Y . (B) Western blot confirming that AR forms aggregates in the presence of 5-H-Y or cisplatin. (C–E) Cellular thermal shift assay results for AR after treatment with dihydrotestosterone (DHT), KW-365, or 5-H-Y . DMSO, dimethyl sulfoxide (vehicle).

Article Snippet: Post SDS-PAGE, the gel was immersed in Coomassie Brilliant Blue stain (CBB Stain One (Nacalai Tesque)) and incubated overnight.

Techniques: Staining, Western Blot, Thermal Shift Assay

Journal: Frontiers in Microbiology

Article Title: Strawberry Vein Banding Virus Movement Protein P1 Interacts With Light-Harvesting Complex II Type 1 Like of Fragaria vesca to Promote Viral Infection

doi: 10.3389/fmicb.2022.884044

Figure Lengend Snippet: FvLHC II-1L assists SVBV P1 in complementing movement-defective potato virus X (PVX). (A) FvLHC II-1L promotes SVBV P1 to complement movement-defective PVX (PVX ΔP25 -GFP) in N. benthamiana . PVX ΔP25 -GFP was co-infiltrated with pBin438 empty vector, P25, SVBV P1, or SVBV P1 and FvLHC II-1L in N. benthamiana . GFP fluorescence was observed at 3 dpi. Bar scale = 100 μm. (B) Western blot analysis of GFP, SVBV P1, or FvLHC II-1L protein accumulation in infiltrated leaves at 3 dpi. α-GFP, α-SVBV P1, and α-myc antibodies were used to detect the accumulation of GFP, SVBV P1, and FvLHC II-1L, respectively. Coomassie Brilliant Blue-stained RuBisCO was used as the loading control. (C) FvLHC II-1L promotes SVBV P1 to complement PVX ΔP25 -GFP in FvLHC II-1L transgenic N. benthamiana . PVX ΔP25 -GFP was co-expressed with SVBV P1 in empty vector (EV) or FvLHC II-1L transgenic N. benthamiana . GFP fluorescence was observed at 3 dpi. (D) The accumulation of GFP, SVBV P1, and FvLHC II-1L proteins in infiltrated leaves was detected via western blotting at 3 dpi.

Article Snippet: After electrophoresis, the gel was placed in Coomassie Brilliant Blue stain (CBB, BeyoBlue TM Coomassie Blue Super Fast Staining Solution; Beyotime, Shanghai, China), soaked for 1 h, then decolorized with pure water.

Techniques: Virus, Plasmid Preparation, Fluorescence, Western Blot, Staining, Control, Transgenic Assay