β tubulin levels  (Cell Signaling Technology Inc)

Journal: Cancer Cell International

Article Title: Association of Porphyromonas gingivalis -infected oral squamous cell carcinoma cell-secreted exosomal miR-3648-1-p5 with tumor progression

doi: 10.1186/s12935-026-04230-5

Figure Lengend Snippet: Verification of exosomes released from Ex, Vp.Ex, and Pg.Ex. (a) Transmission electron microscopy images of Ex, Vp.Ex, and Pg.Ex. Scale bar, 100 nm. (b) Nanoparticle Tracking Analyzer concentration and size in Ex, Vp.Ex, and Pg.Ex mice ( n = 4). (c) Western blot analysis of exosomal markers (CD9, CD63, and TSG101) in Ex, Vp.Ex, and Pg.Ex, with β-tubulin used as the internal control. (d) Experimental design of exosome tracking. SCC25 cells (5 × 10 4 ) are co-cultured with PKH26-labeled Pg.Ex (15 µg mL − 1 ) for different periods. (e) Confocal microscope images of SCC25 cell uptake of fluorescent staining. Scale bar, 10 μm. Data are presented as mean (SD) in b) and c) . * P < 0.05; ** P < 0.01. Data in b) are analyzed using one-way ANOVA

Article Snippet: Horseradish peroxidase-conjugated anti-rabbit antibodies (1:5,000; Beyotime, Shanghai, China) were used as secondary antibodies. β-Tubulin levels were determined using specific antibodies (1:1,000; Cell Signaling Technology, Boston, MA, USA) and served as a loading control.

Techniques: Transmission Assay, Electron Microscopy, Concentration Assay, Western Blot, Control, Cell Culture, Labeling, Microscopy, Staining

Journal: PLOS Genetics

Article Title: Decreased Hsp90 activity protects against TDP-43 neurotoxicity in a C . elegans model of amyotrophic lateral sclerosis

doi: 10.1371/journal.pgen.1011518

Figure Lengend Snippet: A. Radial locomotion assays were used to measure motor function following temperature-based inactivation of HSP-90. Animals were shifted to the restrictive temperature at L4 stage and assessed after 24 hours of HSP-90 inactivation. Animals expressing fALS TDP-43 (M337V) combined with the hsp-90(p673) mutation show an improvement in their motility. Error bars represent Mean with 95% confidence interval (CI): N = 3 independent experimental replicates; total n>100. Statistical significance as determined using Student’s t-test. (**** p<0.0001). B. hsp-90(p673) animals did not move significantly differently from wildtype (N2) animals, which were used as non-transgenic (non-Tg) controls. N = 3 independent experimental replicates; total n>100. Statistical significance as determined using Student’s t-test. (ns = not significant). C-D. Radial locomotion assays were used to measure motor function following pharmacological inhibition of HSP-90. Treatment with the Hsp90 inhibitor 17-AAG improves motor dysfunction caused by the expression of mutant hTDP-43 in C . elegans . Two independent transgenic strains, expressing either TDP-43 (M337V) (C) or TDP-43 (A315T) (D) exhibit improved motility after 48h of 17-AAG treatment (7.5 μM) when scored at 30 minutes, but this effect diminishes or is lost by 60 minutes or 24 hours post-removal of drug. Error bars represent Mean with 95% CI. N = 4; n>100 for the TDP-43 (M337V) treated worms, and n>100 when using the TDP-43 (A315T) strain. Statistical significance as determined using a Mixed-effects analysis (Bonferroni’s multiple comparisons test post hoc test). (*** p<0.001; *p<0.05). E-F . Control worms showed no difference in their motility assessment after 48h of treatment with 17-AAG (7.5 μM). Results shown combined data from multiple experiments. Error bars represent Mean with 95% CI. N = 3; n>100. Statistical significance was determined using a Mixed-effects analysis (Bonferroni’s multiple comparisons test post hoc test). G. Radial locomotion assays were used to measure motor function in older TDP-43 (M337V) at day 3, 5, or 7 of adulthood following a 24 hour temperature-based inactivation of HSP-90. TDP-43 (M337V) ; hsp-90(p673) animals show an improvement in their motility at day 3 of adulthood, but this is lost at days 5 and 7. Error bars represent Mean with 95% CI: N = 3 independent experimental replicates; total n>65. Statistical significance as determined using Student’s t-test. (**** p<0.0001).

Article Snippet: C . elegans β-tubulin levels were measured using monoclonal antibody E7 (Developmental Studies Hybridoma Bank, 1:5000) as a loading control.

Techniques: Expressing, Mutagenesis, Transgenic Assay, Inhibition, Control

Journal: PLOS Genetics

Article Title: Decreased Hsp90 activity protects against TDP-43 neurotoxicity in a C . elegans model of amyotrophic lateral sclerosis

doi: 10.1371/journal.pgen.1011518

Figure Lengend Snippet: Hsp90 mutation or inhibition decreases accumulation of phosphorylated TDP-43 A. Representative immunoblots for total and phosphorylated TDP-43 in C . elegans samples . B-C. Mutant hsp-90 significantly decreases the levels of total and phosphorylated species of TDP-43 (pTDP-43). Error bars represent SEM: N = 4. Statistical significance as determined using unpaired t-test. (*p<0.05, **p< 0.01) D. The ratio of phosphorylated TDP-43 to total TDP-43 (pTDP-43/ total TDP-43) is unchanged in TDP43 (M337V) versus TDP43 (M337V) ; hsp-90(p673) . E. Representative immunoblots for endogenous total and phosphorylated TDP-43 in HEK293T cells pre-treated with increasing amounts of the Hsp90 inhibitor 17-AAG and exposed to Ethacrynic acid 150μM. F-G. Quantification of replicate immunoblots shows no significant differences in total TDP-43 protein levels but a trend towards a reduction in the levels of pTDP-43 in a dose-dependent manner, reaching statistical significance at the highest dose tested, 10μM. Results combined data from multiple experiments. Error bars represent SEM. N = 3. Statistical significance was determined using the One-way ANOVA test (Dunnett’s multiple comparisons post hoc test) (* p<0.05).

Article Snippet: C . elegans β-tubulin levels were measured using monoclonal antibody E7 (Developmental Studies Hybridoma Bank, 1:5000) as a loading control.

Techniques: Mutagenesis, Inhibition, Western Blot

Journal: PLOS Genetics

Article Title: Decreased Hsp90 activity protects against TDP-43 neurotoxicity in a C . elegans model of amyotrophic lateral sclerosis

doi: 10.1371/journal.pgen.1011518

Figure Lengend Snippet: A. Representative photomicrographs of proteasome reporter rpt-3p :: GFP near the tail of non-Tg, hsp-90(p673) , TDP43 (M337V) , and TDP43 (M337V) ; hsp-90(p673) animals. Scale bars = 10μm. B. TDP-43 (M337V) expressing C . elegans increase rpt-3p :: GFP reporter expression, but hsp-90 mutation does not affect reporter expression either independently or in combination with TDP-43 (M337V) . Error bars represent SEM: N = 3, n = 25–31. Statistical significance as determined using the non-parametric Kruskal-Wallis test (Dunn’s multiple comparisons post hoc test).

Article Snippet: C . elegans β-tubulin levels were measured using monoclonal antibody E7 (Developmental Studies Hybridoma Bank, 1:5000) as a loading control.

Techniques: Expressing, Mutagenesis