Journal: PLOS One

Article Title: Voiding symptoms aggravate with decreasing stromal/epithelial ratio and increasing glandular-epithelial content in patients undergoing laser enucleation for benign prostatic hyperplasia, independently from prostate size

doi: 10.1371/journal.pone.0345306

Figure Lengend Snippet: Laser-enucleated prostate tissues were subjected to detection of calponin-1 (CNN1) and keratin-19 (KRT19) mRNA expression by RT-PCR (a, b) , and of calponin and pan-cytokeratin protein expression by Western blot analyses (c) . Shown are all single values from RT-PCR in (a) , which were compared between catheterized (n = 38) and non-catheterized patients (n = 61) in (b) (CNN1 compared by unpaired, two-tailed t-test; KRT19 and CNN1/KRT19 compared by Mann Whitney test; all insignificant). Representative Western blots are shown in (c) . All three blots are from the same sample set. Numbers on the left indicate positions of molecular weight markers. Data for prostate volume, IPSS and Q max from the corresponding patients are indicated on the top (n.a., information not available).

Article Snippet: Subsequently, membranes were washed three times (each time for 5 min) with PBS containing 0.1% Tween 20 (PBS-T), followed by incubation with mouse monoclonal anti pan-cytokeratin (sc-8018), mouse monoclonal anti calponin 1/2/3 (sc-136987), or mouse monoclonal anti-β-actin antibody (sc-47778) (Santa Cruz Biotechnology, Santa Cruz, CA, USA) for 90 min. Primary antibodies were diluted 1:500 in PBS-T containing 5% milk powder.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Two Tailed Test, MANN-WHITNEY, Molecular Weight

Journal: PLOS One

Article Title: Voiding symptoms aggravate with decreasing stromal/epithelial ratio and increasing glandular-epithelial content in patients undergoing laser enucleation for benign prostatic hyperplasia, independently from prostate size

doi: 10.1371/journal.pone.0345306

Figure Lengend Snippet: Intensities of bands from 37-50 kDa in pan-cytokeratin expression (a) , and matching the expected molecular weight of calponin (33 kDa) (b) were referred to band intensities of β-actin in the same samples, and subjected to Pearson correlation analyses with prostate volume, IPSS and Q max in these patients.

Article Snippet: Subsequently, membranes were washed three times (each time for 5 min) with PBS containing 0.1% Tween 20 (PBS-T), followed by incubation with mouse monoclonal anti pan-cytokeratin (sc-8018), mouse monoclonal anti calponin 1/2/3 (sc-136987), or mouse monoclonal anti-β-actin antibody (sc-47778) (Santa Cruz Biotechnology, Santa Cruz, CA, USA) for 90 min. Primary antibodies were diluted 1:500 in PBS-T containing 5% milk powder.

Techniques: Expressing, Molecular Weight

Journal: bioRxiv

Article Title: Fibroblasts neurotrophin signaling sustains pathological vascular maturation in rheumatoid arthritis

doi: 10.64898/2026.03.12.711120

Figure Lengend Snippet: (A-C) Bar plots showing differential regulation of pericyte markers (RGS5, ABCC9), VSMC markers (CNN1, MYH11), and pan-mural cell markers (ACTA2, SMTN, TAGLN) in fibroblasts treated with (A) NGF, (B) BDNF, or (C) NT3 compared with untreated fibroblasts. Bar heights represent log₂ fold change, and bar color indicates the corresponding P values. (D) Schematic of the collagen gel contraction assay in which synovial fibroblasts were embedded in collagen I gels and treatment (E) Representative images of collagen I gel contraction following treatment with NGF, BDNF, or NT3 (F) Quantification of collagen I gel contraction expressed as percent contraction. Values represent mean ± standard deviation (SD). Individual data points indicate biological replicates. Statistical analysis was performed using one-way ANOVA, with P values shown. (G) Schematic illustrating the synovial organoid/explant experimental design. (H) Representative H&E image of synovial explant culture after 3 days of culture. (I) Whole-mount immunoluorescence images showing preservation of endothelial cells after 3 days in culture, marked by CD31 (magenta), and vascular smooth muscle cells marked by MYH11, surrounding CD31 vascular structures. (J) Whole-mount staining showing expression of neurotrophin receptors NGFR, NTRK1, NTRK2, and NTRK3 (green) around vascularature, together with CD31 (magenta) and DAPI (blue). (K) Synovial organoids embedded in Matrigel and treated with NGF, BDNF, or NT3, followed by fixation and staining for a-smooth muscle actin (aSMA) (brown) to visualize mural cells. All 20X images were cropped at similar scale. Scale bar, 150 µm.

Article Snippet: Following transfer, membranes were blocked for 20 minutes in Everyblot blocking buffer (Bio-Rad # 12010020) and incubated overnight at 4°C with primary antibodies from TrkA and TrkB antibody sampler kit against TrkA, TrkB, p-TrkA/TrkB(Cell Signaling Technology, #4638, 1:500), CNN1 (Proteintech-24855-1-AP), MYH11 (Proteintech-21404-1-AP), NGF (Abcam-ab52918) and GAPDH (Thermo Fisher Scientific, #MA5-15738),or beta-actin (Cell Signaling Technology, #3700).

Techniques: Collagen Gel Contraction Assay, Standard Deviation, Preserving, Staining, Expressing

Journal: bioRxiv

Article Title: Fibroblasts neurotrophin signaling sustains pathological vascular maturation in rheumatoid arthritis

doi: 10.64898/2026.03.12.711120

Figure Lengend Snippet: (A) RNAscope images showing transcript expression of pericyte marker RGS5 (yellow) and VSMC marker MYH11 (red) in fibroblasts treated with different neurotrophins, overlaid with DAPI (blue). Scale bar, 150 µm. All images were acquired at 20x magnification and cropped to the same scale.(B) Quantification of total numbers of RGS5+, MYH11+, and RGS5+/MYH11+ double-positive cells following treatment with the indicated neurotrophins.(C) mRNA expression of the pericyte marker RGS5 in fibroblasts transfected with siRNAs targeting NGFR or NTRK1, in the presence or absence of NGF stimulation.(D) mRNA expression of the VSMC marker MYH11 in fibroblasts transfected with siRNAs targeting NTRK2 or NTRK3, in the presence or absence of BDNF or NT3 stimulation, respectively.(E-F) mRNA expression of the mural cell marker CNN1 (calponin) in fibroblasts stimulated with NGF (E) in the presence or absence of a TRKA inhibitor, or stimulated with BDNF (F) in the presence or absence of an NTRK2 inhibitor.

Article Snippet: Following transfer, membranes were blocked for 20 minutes in Everyblot blocking buffer (Bio-Rad # 12010020) and incubated overnight at 4°C with primary antibodies from TrkA and TrkB antibody sampler kit against TrkA, TrkB, p-TrkA/TrkB(Cell Signaling Technology, #4638, 1:500), CNN1 (Proteintech-24855-1-AP), MYH11 (Proteintech-21404-1-AP), NGF (Abcam-ab52918) and GAPDH (Thermo Fisher Scientific, #MA5-15738),or beta-actin (Cell Signaling Technology, #3700).

Techniques: RNAscope, Expressing, Marker, Transfection