hfabtm rhodamine anti tubulin antibody  (Bio-Rad)

Journal: Frontiers in Pharmacology

Article Title: Soluble epoxide hydrolase inhibition restores pro-resolving lipid mediators and reduces inflammation in localized provoked vulvodynia

doi: 10.3389/fphar.2026.1741914

Figure Lengend Snippet: sEH is involved in the LPV inflammatory process. (A) Illustration of punch biopsy collection from LPV cases and healthy controls during surgery. Light touch to the vulvar vestibule (Vest) is immensely painful, whereas the adjacent external vulva (Vulv) is not painful to touch. (B) RT-qPCR data showing sEH (EPHX2) mRNA expression, normalized to 18S rRNA, in LPV and control fibroblasts (n = 6 cases and 6 controls). (C) Western blot data showing sEH (EPHX2) protein expression, normalized to β-tubulin, in LPV and control fibroblasts (n = 3 cases and 3 controls). (D) sEH enzyme activity is significantly elevated in the LPV vest compared to LPV vulv, control vest, and control vulv (n = 3 cases and 3 controls). The data was collected in technical triplicates and are represented as mean ± SEM, one-way ANOVA + Tukey’s post hoc test, *P < 0.05, ns = not significant. (E) siRNA knockdown with sEH siRNA [5 µg] in vulvar and vestibular fibroblasts derived from 3 LPV cases was confirmed through Western blotting. (F) Transfection of sEH siRNA in LPV case vestibular and vulvar fibroblasts, but not control siRNA, reduced IL-6 and PGE2 production upon IL-1β [500 pg/mL] stimulation (n = 3 cases). The data was collected in six technical replicates and are represented as mean ± SEM, two-way ANOVA + Tukey’s post hoc test, *P < 0.05.

Article Snippet: Protein blots were probed with EPHX2 rabbit polyclonal antibody (Abclonal, Woburn, MA, Cat: A1885) at a dilution of 1:1,000, hFAB Rhodamine Anti-Tubulin Primary Antibody (Bio-Rad, Cat: #12004166) at a dilution of 1:5,000, and peroxidase AffiniPure goat anti-rabbit IgG (H + L) (Jackson ImmunoResearch, West Grove, PA, Cat: 111-035-003) at a dilution of 1:4,000.

Techniques: Quantitative RT-PCR, Expressing, Control, Western Blot, Activity Assay, Knockdown, Derivative Assay, Transfection

Journal: Nucleic Acids Research

Article Title: NUP98 regulates orthoflavivirus replication through interaction with vRNA and can be targeted for antiviral purposes

doi: 10.1093/nar/gkag027

Figure Lengend Snippet: BOP1 peptide inhibits orthoflavivirus replication. ( A, B ) CLIP assay of A549 cells transfected with 3xFLAG-NUP98–CTD, untransduced or transduced with lenBOP1 or lenBOP1 Mut infected with TBEV (MOI 0.1) 24 h. Immunoprecipitation performed using FLAG-Trap (triangle) or control beads (square). WB performed using anti-FLAG to detect 3xFLAG-NUP98–CTD, anti-GFP to detect transduced peptides and anti-tubulin as a loading control ( A ). CLIP assay RNA values were normalized to input vRNA and control beads precipitated samples for each condition ( B ). Bars represent mean + SD. n = 6. Significance calculated using one-way ANOVA with Tukey’s multiple comparison test. ns, not significant; ( C ) A549 cells transduced with lentiviruses expressing lenBOP1 or lenBOP1 Mut peptide infected with TBEV MOI 0.1 24 h analyzed by RT-qPCR. vRNA normalized to infection of untransduced control cells. n = 6. Bars represent means + SD. Significance calculated using one-way ANOVA with Tukey’s multiple comparison test. ( D ) A549 cells transduced with lentiviruses expressing lenBOP1 or lenBOP1 Mut peptide infected with different orthoflaviviruses MOI 0.1 (DENV2 MOI 1) 24 h. Infection normalized to infection using transduced lenBOP1 Mut peptide for each virus. n = 6. Bars represent means + SD. Significance calculated with multiple t -test with the Holm–Šídák method for correction. ( E ) Viability assay on A549 cells treated with cpBOP1 or cpBOP1 Mut peptide in different concentrations. n = 3. Dots ± SD fitted with a nonlinear regression curve. Dotted line at 100%. ( F ) A549 cells treated with cpBOP1 or cpBOP1 Mut peptide in different concentrations and infected with TBEV MOI 0.1 for 24 h analyzed using RT-qPCR. Values normalized to infection in samples treated with lowest concentration of control peptide. Dots represents means ± SD fitted with a nonlinear regression curve. The dotted line indicates level of infection in lowest control peptide treated TBEV infected sample. ( G ) Time of addition experiment on A549 cells infected with TBEV MOI 0.03 24 h treated with cpBOP1 or cpBOP1 Mut peptide −2, 1, 3, and 5 hpi and analyzed using RT-qPCR. Samples normalized to no peptide control samples. n = 6. Bars represent means + SD. Statistical analysis performed using two-way ANOVA with Sidak’s multiple comparisons test. ( H ) RT-qPCR replication assay quantifying amount of vRNA in A549 cells untreated or treated with cpBOP1 and cpBOP1 Mut peptide at 8 h post TBEV infection MOI 1. Samples were normalized to input which was collected 2 h post TBEV infection. Bars represent mean + SD. n = 9. Statistical analysis performed using one-way ANOVA with Turkey comparisons test. ( I–K ) Immunofluorescence analysis of A549 cells expressing lenBOP1 Mut or lenBOP1 transfected with TBEV replicon 48 h. anti-NS3 (red), anti-dsRNA (grays), eGFP (green), and DAPI (blue). Scale bars represent 10 µm. Quantification of mean intensity of NS3 ( J ) and dsRNA ( K ) stain in TBEV replicon transfected cells. Bars represent means + SD. n = 30. Statistical analysis performed using unpaired t -test. * < 0.05, ** < 0.01; *** < 0.001; **** < 0.0001.

Article Snippet: Anti-Tubulin hFAB Rhodamine , , , 1:4000 (WB) , BioRad (12004166).

Techniques: Transfection, Transduction, Infection, Immunoprecipitation, Control, Comparison, Expressing, Quantitative RT-PCR, Virus, Viability Assay, Concentration Assay, Immunofluorescence, Staining