Journal: iScience

Article Title: Guanylate-binding protein 1 modulates proteasomal machinery in ovarian cancer

doi: 10.1016/j.isci.2023.108292

Figure Lengend Snippet: GBP1 influences chemotherapy response (A–D) 100–300 transduced OC cells/well were plated in 12 well tissue culture plates followed by treatment with paclitaxel (ID8-15 nM and OVCAR8-7.5 nM) after 24 h and allowed to grow for 7–10 days. Colonies were stained using crystal violet and confluency was measured. Data are shown as mean ± SD of triplicate samples. Significant differences compared to respective controls were assessed by Student’s t test. (E) Immunoblotting of total ubiquitinated proteins in transduced OC cells treated with vehicle (DMSO) (C) or paclitaxel (ID8-15 nM and OVCAR8-7.5 nM) for 24 h. (F–I) Kaplan-Meier plots (KMplot) for the probability of survival of ovarian cancer patients with GBP1-high (high) and GBP1-low (low) expression (mRNA gene chip data, Affy ID: 202269_x_at). KMplots were plotted using KM-plotter. (F) KMplot (n = 1435) of ovarian cancer patients with high and low GBP1 expression. The plot includes patients of all the stages (1–4) of ovarian cancer. (G) KMplot (n = 163) of ovarian cancer patients with disease at stages 1 and 2 with high and low GBP1 expression. (H) KMplot (n = 73) of ovarian cancer patients with disease at stages 1 and 2 and treated with Taxol with high and low GBP1 expression. (I) KMplot (n = 94) of ovarian cancer patients with disease at stages 1 and 2 and treated with platinum agents with high and low GBP1 expression.

Article Snippet: Paclitaxel (Cat. No. # 10461100) was purchased from Cayman Chemical, USA, and stock solutions were prepared in DMSO (MP Biomedicals, #196055) and stored at -20°C.

Techniques: Staining, Western Blot, Expressing

Journal: iScience

Article Title: Guanylate-binding protein 1 modulates proteasomal machinery in ovarian cancer

doi: 10.1016/j.isci.2023.108292

Figure Lengend Snippet:

Article Snippet: Paclitaxel (Cat. No. # 10461100) was purchased from Cayman Chemical, USA, and stock solutions were prepared in DMSO (MP Biomedicals, #196055) and stored at -20°C.

Techniques: Recombinant, Over Expression, Plasmid Preparation, shRNA, Control, Lysis, Activity Assay, Mass Spectrometry, Software

Journal: Experimental neurology

Article Title: Independent evaluation of the anatomical and behavioral effects of Taxol in rat models of spinal cord injury

doi: 10.1016/j.expneurol.2014.06.020

Figure Lengend Snippet: Custom designed catheters for intrathecal delivery of cremaphor vehicle and/or Taxol. A rat intrathecal catheter (Alzet #0007740) was modified by inserting a segment of silastic tubing at the junction between the osmotic pump and the remaining segment of intrathecal catheter. (1) Briefly, intrathecal catheters, comprised of a thin Teflon-coated wire stylet (A), an external catheter segment (B), a connector segment (C) and the intrathecal segment (D) were cut to a length of ~50 mm. Three separate pieces of silastic tubing (Dow Corning; #508-004) also were prepared for each catheter; two cuffs (4mm and 2mm) and a single 20 mm segment (E, F, G). (2) The external segment of catheter is cut to a length of ~8mm (B) then is inserted along with the stylet (A) into one end of the 20 mm segment of silastic tubing (E) until it abuts the connector (C) just proximal to the distal catheter segment (D). (3) A 4 mm silastic cuff (F) was slipped over this junction (H) then the external catheter segment was connected to the metal hub of the osmotic pump. (4) The remaining 2 mm silastic cuff (G) was placed mid way over the distal 20 mm segment of silastic tubing where it serves as an anchor point for securing sutures on each side of the cuff. (5) Finally, a few drops of Histoacryl® (TissueSeal, LLC; #TS1050071FP; Ann Arbor, MI) was applied to all cuffs (F,G) and at the interface between the long segment of silastic tubing (E) and the pump connector. This seals all connections and prevents leakage.

Article Snippet: Drug preparation Taxol (Paclitaxel; cat #9600 LC Laboratories; Woburn, MA) was purchased just prior to the start of the first experiments and stored as a lyophilized powder at −20°C until use.

Techniques: Modification

Journal: Experimental neurology

Article Title: Independent evaluation of the anatomical and behavioral effects of Taxol in rat models of spinal cord injury

doi: 10.1016/j.expneurol.2014.06.020

Figure Lengend Snippet: Expanded anatomical analysis of spinal cord hemisection lesion after 7-day infusion with vehicle or Taxol. (A) Lesion centers from two vehicle and two Taxol specimens stained with EC/CV. Note the extended cap of connective tissue (black arrows) that is continuous with the lesion matrix in tissues from vehicle treatment group. These are absent in specimens from the Taxol-treated group (scale = 500 μm). (B) Taxol has no effect on the dorsal-ventral (DV) lesion length or total volume as measured from EC/CV stained sections. (C) Taxol reduces deposition of laminin, collagen IV and fibronectin (scale = 400 μm) and alters CS56 expression within the lesion (scale = 200 μm). Note how the diffuse pattern of fibronectin labeling in vehicle treated (white arrow) contrasts with the dense fibrillar staining pattern in Taxol-treated specimens (double white arrows). (D) GFAP- immunoreactivity lines the lesion border in both vehicle and taxol treated specimens. (F) Immunofluorescence intensity was measured in a constant region of interest (0.04 mm2 sample box) positioned at the rostral lesion border (scale = 200 μm). Both fibronectin and GFAP labeling were increased at this site after Taxol infusion. (F) Qualitative analyses reveal fewer neurofilament-positive axons and NG2+ profiles (asterisks) in Taxol-treated specimens (scale = 100 μm).

Article Snippet: Drug preparation Taxol (Paclitaxel; cat #9600 LC Laboratories; Woburn, MA) was purchased just prior to the start of the first experiments and stored as a lyophilized powder at −20°C until use.

Techniques: Staining, Expressing, Labeling, Immunofluorescence

Journal: Experimental neurology

Article Title: Independent evaluation of the anatomical and behavioral effects of Taxol in rat models of spinal cord injury

doi: 10.1016/j.expneurol.2014.06.020

Figure Lengend Snippet: Taxol infusion reduces fibrotic scarring in acute dorsal spinal hemisection lesions (cf. Fig. 1 in Hellal et al). (A) Schematic illustrating location (box) where images were captured for analysis. (B,C) Mid-sagittal sections through boxed region in (A) showing distinct staining patterns for laminin (green) and GFAP (red) within the lesion site following vehicle or Taxol infusion. (D) Taxol decreased laminin and fibronectin staining in the lesion, expressed as % of the average value from vehicle group (2- tailed unpaired t-test, ***p<0.001; fibronectin staining t-test with Welch's correction **p<0.01). (E) GFAP+ stain at the lesion border was scored using the 0-3 scale used by Hellal. Two-way ANOVA with repeated measures (distance) revealed significant effects of distance and subject matching (p<0.0001), but no effect of treatment. (F) Taxol did not affect lesion size (2 tailed t-test, p=0.225). (n=14/group for all measures; Scale in A =200 μ)

Article Snippet: Drug preparation Taxol (Paclitaxel; cat #9600 LC Laboratories; Woburn, MA) was purchased just prior to the start of the first experiments and stored as a lyophilized powder at −20°C until use.

Techniques: Staining

Journal: Experimental neurology

Article Title: Independent evaluation of the anatomical and behavioral effects of Taxol in rat models of spinal cord injury

doi: 10.1016/j.expneurol.2014.06.020

Figure Lengend Snippet: Taxol infusion does not improve locomotor function after contusive SCI. Recovery of motor function as measured on an elevated ladder (A) was not different between Taxol and vehicle-treated rats. Spontaneous recovery of locomotor function, as measured in the open-field using Basso-Beattie-Bresnahan (BBB) locomotor rating scale (B) or BBB sub-scoring (C) also was unchanged by Taxol.

Article Snippet: Drug preparation Taxol (Paclitaxel; cat #9600 LC Laboratories; Woburn, MA) was purchased just prior to the start of the first experiments and stored as a lyophilized powder at −20°C until use.

Techniques:

Journal: Experimental neurology

Article Title: Independent evaluation of the anatomical and behavioral effects of Taxol in rat models of spinal cord injury

doi: 10.1016/j.expneurol.2014.06.020

Figure Lengend Snippet: Taxol infusion does not affect contusion lesion size or number of serotonin (5HT+) axons but reduces intralesional matrix deposition. (A) Representative photomicrographs of EC/CV-stained sagittal sections 56 days after spinal contusion injury. Sections were cut through the lesion epicenter or 250 μm to the right or left of center (L= left of center, C=center, R= right of center; scale = 200 μm). (B) Both groups have similar mean lesion volume; however, Taxol reduced intralesional accumulation of tissue matrix (unpaired t-test; n=9-11/group; *** P<0.001). (C) Sagittal sections through the center of the contusion lesion stained with anti-5HT. Robust 5HT-positive axon labeling is evident in the dorsal region caudal to the lesion in both groups (boxed region enlarged in right panels, axons noted with arrowheads). Scale in left panels = 200 μm; right panels = 50 μm. (D) Taxol did not increase the number of 5-HT axons in the contused spinal cord. (E) The lesion site is filled with laminin-enriched ECM in vehicle-treated but not in Taxol-treated sections. Sections in (E) are cut through the lesion center. Scale = 200 μm. (F) The laminin-positive area within the lesion borders is reduced by Taxol (unpaired t-test; N=9-11/group; *** p<0.001).

Article Snippet: Drug preparation Taxol (Paclitaxel; cat #9600 LC Laboratories; Woburn, MA) was purchased just prior to the start of the first experiments and stored as a lyophilized powder at −20°C until use.

Techniques: Staining, Labeling

Journal: Experimental neurology

Article Title: Independent evaluation of the anatomical and behavioral effects of Taxol in rat models of spinal cord injury

doi: 10.1016/j.expneurol.2014.06.020

Figure Lengend Snippet: Taxol infusion reduces CSPG accumulation but does not support axon growth/sprouting after contusive SCI. (A) Taxol infusion reduces the area of CS-56 labeling at the lesion borders, while dense staining is evident along the dorsal meningeal surface (*). Scale = 200 μm. (B) CS-56 immunoreactivity area as a proportion of the region of interest at the borders. 2-way ANOVA with repeated measures revealed significant overall effect of Taxol (p= 0.02) with a difference in staining at the rostral lesion border by post-hoc analysis (p<0.05; n=9-11/group). (C) Double staining with GFAP (green) and neurofilament (NF) (red) antibodies reveals dense penetration by axons into the laminin-rich lesion site in vehicle-treated subjects (see also Fig. 6E). Boxed area is enlarged to the right. Conversely, few axons penetrate the acellular void that defines the lesion site in Taxol treated specimens. Scale in left panels = 200 μm, scale in enlarged panels = 100 μm.

Article Snippet: Drug preparation Taxol (Paclitaxel; cat #9600 LC Laboratories; Woburn, MA) was purchased just prior to the start of the first experiments and stored as a lyophilized powder at −20°C until use.

Techniques: Labeling, Staining, Double Staining