septin 9  (Proteintech)

Journal: European journal of cell biology

Article Title: Septin-dependent defense mechanisms against Pseudomonas aeruginosa are stalled in cystic fibrosis bronchial epithelial cells.

doi: 10.1016/j.ejcb.2024.151416

Figure Lengend Snippet: Fig. 2. SEPT7 mRNA expression analysis using qPCR (A) and SEPT7 (B, C), SEPT2 (B, D), SEPT6 (B, D), SEPT9 (B, D), SEPT11 (B, D) protein expression using western blot in BEAS-2B after 72 h transfection of 5 nM control siRNA (siCTL, white circles) or SEPT7 siRNA (blue circles) (n=4, *p<0.05, Mann–Whitney test). (E) Amount of intracellular P. aeruginosa (Pa) (CFU/mL) in BEAS-2B cells transfected for 72 h with 5 nM of siCTL or SEPT7 siRNA and then infected for 30 min (orange circles, n=6), 1 h (purple circles, n=5) and 3 h (red circles, n=10) with Pa (MOI=5) [(*p<0.05, analysis of variance (ANOVA), Bonferroni correction)]. (F) IL-6 production by BEAS-2B cells transfected for 72 h with 5 nM of siCTL or SEPT7 siRNA in the non-infected (-, white circles) condition and after 3 h of Pa (red cir cles) infection (MOI=5) [(n= 9; *p<0.05, **p<0.01, ****p<0.0001 analysis of variance (ANOVA), Bonferroni correction)].

Article Snippet: The following SEPT antibodies have already been validated (Pfanzelter et al., 2018): SEPT2 (60075–1, 1/20 000 Proteintech, Manchester, UK), SEPT6 (HPA005665, 1/1000, Atlas Antibody, Bromma, Sweden), SEPT7 (18991; 1/1000, IBL, Minneapolis, MN, USA), SEPT9 (A302–353A-M, 1/1000 Bethyl Laboratories, Montgomery, Texas), and SEPT11 (A304–1761-M, 1/1000, Bethyl laboratories).

Techniques: Expressing, Western Blot, Transfection, Control, MANN-WHITNEY, Infection

Journal: The Journal of Cell Biology

Article Title: A septin GTPase scaffold of dynein–dynactin motors triggers retrograde lysosome transport

doi: 10.1083/jcb.202005219

Figure Lengend Snippet: COS-7 cells were cotransfected with LAMP1-mGFP and SEPT9_i1-mCherry under the control of a weak PGK promoter and imaged every 3 s by spinning-disk microscopy. Arrowheads point to SEPT9-positive lysosomes moving retrogradely. SEPT9_i1-mCherry is pseudo-colored in inverted magenta, and LAMP1-mGFP is pseudo-colored in inverted green. Scale bar, 20 µm. Video display rate is 5 frames per second. Related to Fig. 1 .

Article Snippet: Cells were immunostained with the following antibodies: mouse anti-Lamp1 (1:100; H4A3 clone; Iowa Developmental Studies Hybridoma Bank), rabbit anti-SEPT9 (1:100; 10769-1-AP; Proteintech Group), rabbit anti-SEPT9 (1:200; NBP2-13294; Novus Biologicals), mouse anti-SEPT9 clone 10C10 (1:100; MABE992; Millipore-Sigma), rabbit anti-SEPT2 (N5N; 1:300; gift from Makoto Kinoshita, Nagoya University, Nagoya, Japan), rabbit anti-SEPT7 (1:300; 18991; IBL America), rabbit anti-SEPT6 (1:300; gift from Makoto Kinoshita), mouse anti-GM130 (1:200; 610823; BD Biosciences), mouse anti-DIC antibody clone 74.1 (1:100; MAB1618; Millipore), rabbit anti-LAMTOR4 (1:200; 12284S; Cell Signaling), and mouse anti-EEA1 clone 14/EEA1 (1:100; 610456; BD Biosciences).

Techniques:

Journal: The Journal of Cell Biology

Article Title: A septin GTPase scaffold of dynein–dynactin motors triggers retrograde lysosome transport

doi: 10.1083/jcb.202005219

Figure Lengend Snippet: Rat embryonic hippocampal neurons (DIV4) were cotransfected with rat SEPT9_i1-mCherry and LAMP1-mGFP and imaged with shallow-angle TIRF microscopy every 1 s for 2 min. Rat SEPT9_i1-mCherry is pseudo-colored in magenta, and LAMP1-mGFP is pseudo-colored in green. Video display rate is 15 frames per second. Neuronal cell body (soma) is located to the left to the axonal segment of the video (i.e., particles move retrogradely to the left and anterogradely to the right). Scale bar, 5 µm. Related to Fig. 1 .

Article Snippet: Cells were immunostained with the following antibodies: mouse anti-Lamp1 (1:100; H4A3 clone; Iowa Developmental Studies Hybridoma Bank), rabbit anti-SEPT9 (1:100; 10769-1-AP; Proteintech Group), rabbit anti-SEPT9 (1:200; NBP2-13294; Novus Biologicals), mouse anti-SEPT9 clone 10C10 (1:100; MABE992; Millipore-Sigma), rabbit anti-SEPT2 (N5N; 1:300; gift from Makoto Kinoshita, Nagoya University, Nagoya, Japan), rabbit anti-SEPT7 (1:300; 18991; IBL America), rabbit anti-SEPT6 (1:300; gift from Makoto Kinoshita), mouse anti-GM130 (1:200; 610823; BD Biosciences), mouse anti-DIC antibody clone 74.1 (1:100; MAB1618; Millipore), rabbit anti-LAMTOR4 (1:200; 12284S; Cell Signaling), and mouse anti-EEA1 clone 14/EEA1 (1:100; 610456; BD Biosciences).

Techniques: