Journal: Renal Failure

Article Title: ABT-263 exerts a protective effect on upper urinary tract damage by alleviating neurogenic bladder fibrosis

doi: 10.1080/0886022x.2023.2194440

Figure Lengend Snippet: Figure 4. Staining of bladder tissue sections in each group. (a1–e1) represent the general condition of the bladder in the sham group, sham þ ABT-263 group (50 mg/kg), NBF group, NBF þ ABT-263 group (25 mg/kg), and NBF þ ABT-263 group (50 mg/kg), respectively. (a2–e2) represents the matoxylin and eosin (HE) staining of the bladder in each group. (a3–e3) represent Masson staining of each bladder. a4–e4 represents Sirius red staining of the bladder in each group. (f) Statistical histogram of bladder weight in each group, which shows that bladder fibrosis was basically normal in the sham group and sham þ ABT-263 group (50 mg/kg), the most serious in the NBF group, slightly improved in the NBF þ ABT-263 group (25 mg/kg), and greatly improved in the NBF þ ABT-263 group (50 mg/kg). (g) Statistics of the bladder HE staining score in each group histogram, which showed that bladder inflammation was basically normal in the sham and sham þ ABT-263 (50 mg/kg) groups, the most serious in the NBF group, slightly improved in the NBF þ ABT-263 group (25 mg/kg), and greatly improved in the NBF þ ABT-263 group (50 mg/kg). (h) Statistical histogram of bladder fibrosis area in each group, which shows that bladder fibrosis was basically normal in the sham and sham þ ABT-263 (50 mg/kg) groups, the most serious in the NBF group, slightly improved in the NBF þ ABT-263 group (25 mg/kg), and greatly improved in the NBF þ ABT-263 group (50 mg/kg). j: Statistical histogram of the Col3/Col1 ratios for each group. HE Scale bar ¼ 200 lm. Masson Scale bar ¼ 50 lm. Sirius red Scalebar ¼ 100 lm. Data are expressed as the mean ± SEM; ns p > 0.5, p < 0.01, p < 0.001, p < 0. 0001. All experiments were repeated three times.

Article Snippet: SDS-PAGE was performed, and the membrane was transferred, blocked with 5% milk on a shaker for 1 h, and incubated with the following primary antibodies: col1 (SC-393573), col3 (SC-271249), FN(SC8422)(Santa Cruz Biotechnology, Dallas, TX, USA); TGF-b1 (21898-1-AP; Proteintech, Wuhan, China); a-SMA (CST-48938S); BCL-xL (CST-2764S, CST, Danvers, MA, USA); GAPDH, (CST-8884, CST, Danvers, MA, USA).

Techniques: Staining

Journal: Renal Failure

Article Title: ABT-263 exerts a protective effect on upper urinary tract damage by alleviating neurogenic bladder fibrosis

doi: 10.1080/0886022x.2023.2194440

Figure Lengend Snippet: Figure 5. WB and qPCR results of col1, col3, and FN in the bladder tissues of different groups. (a–c) Statistical histogram of the relative ratio of col1, col3, and Fn mRNA in bladder tissue in the sham, sham þ ABT-263 (50 mg/kg), NBF group, NBF þ ABT-263 (25 mg/kg), NBF þ ABT-263 (50 mg/kg) groups; d: WB in bladder tissue in sham, sham þ ABT-263 (50 mg/kg), NBF, NBF þ ABT-263 (25 mg/kg), and NBF þ ABT-263 (50 mg/kg) groups, with GADPH as the internal reference protein. (e–g) Relative expression of col1, col3, and FN protein in bladder tissue in sham, sham þ ABT-263 (50 mg/kg), NBF, NBF þ ABT-263 (25 mg/kg), and NBF þ ABT-263 (50 mg/kg) groups. The histogram of ratio statistics shows that bladder fibrosis were basically normal in the sham and sham þ ABT-263 (50 mg/kg) groups, the most serious in the NBF group, slightly improved in the NBF þ ABT-263 group (25 mg/kg), and greatly improved in the NBF þ ABT-263 group (50 mg/kg). Data are expressed as the mean ± SEM; ns p > 0.5, p < 0.01, p < 0.001, p < 0.0001. All experiments were repeated three times.

Article Snippet: SDS-PAGE was performed, and the membrane was transferred, blocked with 5% milk on a shaker for 1 h, and incubated with the following primary antibodies: col1 (SC-393573), col3 (SC-271249), FN(SC8422)(Santa Cruz Biotechnology, Dallas, TX, USA); TGF-b1 (21898-1-AP; Proteintech, Wuhan, China); a-SMA (CST-48938S); BCL-xL (CST-2764S, CST, Danvers, MA, USA); GAPDH, (CST-8884, CST, Danvers, MA, USA).

Techniques: Expressing

Journal: Cell reports methods

Article Title: Human MAPT knockin mouse models of frontotemporal dementia for the neurodegenerative research community.

doi: 10.1016/j.crmeth.2025.101024

Figure Lengend Snippet: Figure 3. Immunohistochemical analysis of triple-mutant MAPT KI mouse lines (A) Epitope maps of anti-tau antibodies used for this study. (B) Immunohistochemical analysis of HY of male MAPT, MAPTS305N;Int10+3;S320F, and MAPTP301S;Int10+3;S320F mice at the age of 6 months using CP13, AT8, PHF-1, TOC1, T22, and MC1 antibodies. Scale bars: 50 mm; n = 4 (2 males and 2 females). (C) Colocalization of AT8 signals with MAP2 (a marker for somata and dendrites, top) or KIF5A (a marker for somata and axons, bottom) in the cortical region of 12-month-old MAPTP301S; Int10+3; S320F mice. Scale bars: 5 mm. Pink, white, and red arrows indicate dendrite, soma, and axon, respectively. See also Figure S3.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies CP13 Kindly provided by Peter Davies RRID:AB_2314223 AT8 Thermo Fisher Scientific Cat# MN1020; RRID:AB_223647 PHF1 Kindly provided by Peter Davies RRID:AB_2315150 TOC1 Novus Cat# NBP3-20163; RRID:AB_3608371 T22 Sigma-Aldrich Cat# ABN454; RRID:AB_2888681 MC1 Kindly provided by Peter Davies RRID:AB_2314773 Tau13 Santa Cruz Biotechnology Cat# sc-21796; RRID:AB_628328 Tau5 Thermo Fisher Scientific Cat# AHB0042; RRID:AB_2536235 MAP2 Abcum Cat# ab183830; RRID:AB_2895301 KIF5A Proteintech Cat# 21186-1-AP; RRID:AB_10733125 GA5 (Anti-GFAP) Merck Millipore Cat# MAB360; RRID:AB_11212597 Anti-Iba1 Fujifilm-wako Cat# 013–27691; RRID:AB_2934095 Synaptotagmin Synaptic System Cat# 105-002; RRID:AB_887830 Homer1 Synaptic System Cat# 160-004; RRID:AB_2619855 Synaptophysin PROGEN Cat# 61012 PSD-95 Synaptic System Cat# 124-011; RRID:AB_2619799 N1D (Anti-Ab) In-house (Saido et al. 1994) N/A Anti-b-actin, HRP Proteintech Cat# HRP-60008; RRID:AB_2819183 Anti-Rabbit IgG, HRP Proteintech Cat# SA00001-2; RRID:AB_2722564 Anti-Mouse IgG, HRP Proteintech Cat# SA00001-1; RRID:AB_2722565 Chemicals, peptides, and recombinant proteins 4% paraformaldehyde Nacalai Tesque Cat# 09154-85 RNAiso Plus Takara Bio Cat# 9109 ReverTra Ace TOYOBO Cat# FSQ-301 cOmplete protease inhibitor cocktail Roche Diagnostics Cat# 11697498001 lambda-phosphatase Santa Cruz Biotechnology Cat# sc-200312A ECL prime blocking buffer GE healthcare Cat# RPN418 ECL Select detection reagent GE Healthcare Cat# RPN2235 Tris-EDTA buffer (pH 8.0) (TE buffer) Nacalai Tesque Cat# 06890-54 0.5% blocking reagent powder AKOYA Biosciences Cat# SKU FP1012 VECTASTAIN Elite ABC-HRP Kit Vector Laboratories Cat# PK-6100 DMEM Thermo Fisher Scientific Cat# 41966-029 Lipofectamine 2000 Invitrogen Cat# 11668019 Opti-MEM Gibco Cat# 31985070 Herculase II Fusion DNA Polymerase Agilent Technologies Cat# 600675 mMESSAGE mMACHINE T7 Ultra Transcription kit Thermo Fisher Scientific Cat# AM1345 MEGAshortscript T7 Thermo Fisher Scientific Cat# AM1354 MEGAclear Thermo Fisher Scientific Cat# AM1908 Ex Taq-Polymerase kit Takara Bio Cat# RR001A PhosSTOP Roche Diagnostics Cat# 12352204 ProLong Gold Antifade Mountant Invitrogen Cat# P36934 (Continued on next page) e1 Cell Reports Methods 5, 101024, April 21, 2025

Techniques: Immunohistochemical staining, Mutagenesis, Marker

Journal: NPJ Breast Cancer

Article Title: JAK/STAT1-interferon-ISGylation networks in breast cancer resistance to inhibitors of FOXM1 and CDK4/6

doi: 10.1038/s41523-026-00911-6

Figure Lengend Snippet: A Western protein immunoblots of cell extracts from 73 R, PalboR, and AbemaR cells in their basal condition (Veh treatment) or after 24 h treatment with IFNγ (50 ng/ml), or ruxolitinib (rux, 5 µM) or IFNγ plus rux. Levels corrected for loading control β-actin are shown with the WT Veh level set at 1.0. B In situ cell immunofluorescence for ISG15, STAT1, and ERα in parental WT and resistant cells. For each cell type, immunofluorescence intensities of all cells in 3 fields were individually measured and are shown, along with mean ± SEM. Scale bar is 20 microns. CTCF, corrected total cell fluorescence; CNF, corrected nuclear fluorescence. Statistics are one-way ANOVA followed by Fishers LSD multiple comparisons post hoc test. C Expression of STAT1 and several IFN signature genes ( ISG15, IFI27, IFI44, IFIT1, IFIT3 ) in WT, 73 R, PalboR, and Abema R cells. Cells were treated with vehicle, 50 ng/ml IFNγ, 5 μM rux, or IFNγ plus rux for 24 h. RNA was then extracted from cells and gene expression was monitored by qPCR. Triplicate determinations are shown as dots ± SD. Statistical analysis used the Kruskal-Wallis test B and one-way ANOVA followed by Fisher’s LSD Multiple Comparisons post hoc test C with p-values shown above the brackets. *, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001; ****, p ≤ 0.0001. WT Veh is set at 1.

Article Snippet: Primary antibodies used include STAT1 (RRID:AB_2737027, 1:400 dilution), ISG15 (Cell Signaling Technology, RRID:AB_2126201, 1:100 dilution), and estrogen receptor α (Santa Cruz Biotechnology HC-20, cat. #sc-543, 1:50 dilution).

Techniques: Western Blot, Control, In Situ, Immunofluorescence, Fluorescence, Expressing, Gene Expression

Journal: NPJ Breast Cancer

Article Title: JAK/STAT1-interferon-ISGylation networks in breast cancer resistance to inhibitors of FOXM1 and CDK4/6

doi: 10.1038/s41523-026-00911-6

Figure Lengend Snippet: A Western blots showing free ISG15 protein and ISGylated proteins in these cells. Cells were treated with control vehicle or 50 ng/ml IFNγ for 24 h. Protein extracts were then prepared and examined for ISGylation on Western blots with antibody that detects ISG15 and ISGylated proteins. Protein bands A-E are highlighted. β-actin in the same cell samples was monitored on a separate Western blot and this is shown at the bottom of the figure. Molecular weight markers (in kDa) are shown in the left-most lane. B STAT1 is an ISGylated protein in 73 R, PalboR, and AbemaR cells. Cell extracts were incubated with IgG control antibody or ISG15 antibody and the immunoprecipitated proteins were probed with a STAT1 antibody in the eluate and boiled bead extracts by Western blot. STAT1α (91 kDa) and STAT1β (84 kDa) are observed. Independent immunoprecipitation experiments were conducted twice for all cells. C Primers specific for STAT1α and STAT1β were used to detect these RNAs by qPCR in WT and resistant cells. Cells were incubated with Veh or 50 ng/ml IFNγ for 24 h prior to RNA analysis. Two independent experiments were conducted, and individual triplicate determinations are shown as dots ± SD. Statistical analysis by one-way ANOVA followed by Fisher’s LSD Multiple Comparisons post hoc test with p-values shown above the brackets. *, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001; ****, p ≤ 0.0001. WT Veh is set at 1.

Article Snippet: Primary antibodies used include STAT1 (RRID:AB_2737027, 1:400 dilution), ISG15 (Cell Signaling Technology, RRID:AB_2126201, 1:100 dilution), and estrogen receptor α (Santa Cruz Biotechnology HC-20, cat. #sc-543, 1:50 dilution).

Techniques: Western Blot, Control, Molecular Weight, Incubation, Immunoprecipitation