Journal: Nature Communications

Article Title: Septin 9 induces lipid droplets growth by a phosphatidylinositol-5-phosphate and microtubule-dependent mechanism hijacked by HCV

doi: 10.1038/ncomms12203

Figure Lengend Snippet: ( a ) PIP strip overlay assay: PIP strips were incubated either with glutathion-S-transferase (GST) used as a negative control or with purified septin 9_i3 GST tagged at 0.5 μg ml −1 and analysed with anti GST antibody. LPA, lysophosphatidic acid, LPC, lysophosphocholine, PtdIns, phosphatidylinositol, PtdIns(3)P, PtdIns(4)P, PtdIns(5)P, PtdIns(3,4)P2, PtdIns(3,5)P2, PtdIns(4,5)P2, PtdIns(3,4,5)P3, PA, phosphatidic acid, PS, phosphatidylserine, PE, phosphatidylethanolamine, PC, phosphatidylcholine, S1P, sphingosine 1-phosphate. ( b ) Huh7R cells were treated with cell-permeant PtdIns3P, PtdIns4P, PtdIns5P or PtdIns (4, 5) P2 at 30 μM for 15 min before fixing and staining for septin 9 (green) and LDs (red). The dot squares indicate the zoom area. ( c ) Bar graph shows the LD siz in Huh7R cells treated as described in b . The results were obtained from at least 20 cells for each treatment from three independent experiments. ( d ) Huh7R cells were transfected with IpgD-GFP construct or not (control) for 48 h and stained for LDs (red). Bar graph represents LDs size measured in 15 cells from two independent experiments. Values are means±s.e.m. Student's t -test was used. ** P <0.001, *** P <0.0001. Scale bar, 10 μm.

Article Snippet: Phosphoinositides (PtdIns4P diC8 Cat#P4008, PtdIns5P diC4 Cat#P5004, PtdIns3P diC8 Cat#P3008, PtdIns(4,5)P2 diC8 Cat#P4508) and PIP strip Cat#P-6001 were purchased from Echelon.

Techniques: Stripping Membranes, Overlay Assay, Incubation, Negative Control, Purification, Staining, Transfection, Construct

Journal: Nature Communications

Article Title: Septin 9 induces lipid droplets growth by a phosphatidylinositol-5-phosphate and microtubule-dependent mechanism hijacked by HCV

doi: 10.1038/ncomms12203

Figure Lengend Snippet: ( a ) Septin 9 domain organization: the polybasic domain is located at N-terminal of GTP-binding domain, which is recognized by three motifs G1 (GXXXXGK=G 305 QSGLGK 311 ), G3 (DXXG=D 362 TPG 365 ), G4 (XKXD=A 444 KAD 447 ). Down: septin 9_i1 and septin 9_del1 sequence in the polybasic domain region. The boxed sequence ( 289 RRKAMK 294 ) has been deleted in septin 9_del1. ( b ) Coomassie blue stained SDS–PAGE gel of purified septin 9_i1 and septin 9_del1. ( c ) Immunoblot analysis of purified septin 9_i1 and septin 9_del1. ( d ) PIP strip overlay assay for V5 tag, purified septin 9_i1 and septin 9_del1 performed as in . Lipid-bound V5 fusion proteins were detected with anti-V5 antibody. ( e ) Left: Huh7R cells were transfected either with EV or septin 9_i1 or septin 9_del1 and stained for V5 tag (green) and LDs (red). Right: radial profile plots of the presented cells show LDs intensity distribution in the peripheral and perinuclear regions. ( f ) LDs intensity in the perinuclear and peripheral regions of 36 cells from 5 independent experiments performed as described in e . ( g ) LD size and LD number in 36 cells from 5 independent experiments done as described in e . ( h ) Staining of V5 tag (green) and microtubules (MTs) with β tubulin (red) in Huh7R cells transfected with septin 9_i1 or septin 9 del1. The Pearson's correlation coefficient (Rr) measured in 30 cells from two independent experiments is presented on the merge panels. The right panels are green, red, line profile blots of the pink lines. Values are means±s.e.m. Student's t -test was used. * P <0.05, ** P <0.001, *** P <0.0001. Scale bar, 10 μm.

Article Snippet: Phosphoinositides (PtdIns4P diC8 Cat#P4008, PtdIns5P diC4 Cat#P5004, PtdIns3P diC8 Cat#P3008, PtdIns(4,5)P2 diC8 Cat#P4508) and PIP strip Cat#P-6001 were purchased from Echelon.

Techniques: Binding Assay, Sequencing, Staining, SDS Page, Purification, Western Blot, Stripping Membranes, Overlay Assay, Transfection

Journal: Cell host & microbe

Article Title: The Stringent Response Determines the Ability of a Commensal Bacterium to Survive Starvation and to Persist in the Gut

doi: 10.1016/j.chom.2018.06.002

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Guanosine tetraphosphate, ppGpp , TriLink , Cat#N-6001.

Techniques: Virus, Recombinant, Membrane, Bacteria, Software