Journal: Cancer Research Communications

Article Title: Phase I First-in-Human Study of TRK-950, an IgG1 Antibody Specific to CAPRIN-1, in Patients with Advanced Solid Tumors

doi: 10.1158/2767-9764.CRC-25-0123

Figure Lengend Snippet: Graphs depicting the density of macrophage subsets pre- and posttreatment with TRK-950. Tumor specimens obtained with core-needle biopsies at the screening and C1D22 time points were subjected to multi-immunofluorescence to detect macrophage infiltrates. Quantitative analyses were performed on digital images of CD68 and CD163 staining. A, total CD68 + cells; ( B ) CD68 + /CD163 − subsets; ( C ) CD68 + /CD163 + subsets. Error bars indicate SD. C1D22, cycle 1 day 22; SCR, screening.

Article Snippet: After antigen retrieval (36 minutes at 95°C, pH 8.4), 4-μm-thick sections of tumor specimens obtained from core-needle biopsies at the screening and C1D22 time points were stained with mouse monoclonal anti-human CD68 (IgG2b, clone 298807, R&D Systems) and mouse monoclonal anti-human CD163 (IgG1, clone 10D6, Leica Biosystems) and then incubated with AF647 goat anti-mouse IgG2b and AF488 goat anti-mouse IgG1 (both from Invitrogen).

Techniques: Immunofluorescence, Staining

Journal: Communications Biology

Article Title: Inhibition of inner ear macrophage phagocytosis alleviates cisplatin-induced ototoxicity

doi: 10.1038/s42003-025-08525-7

Figure Lengend Snippet: A Cisplatin treatment timeline for C57BL/6 mice. Created in BioRender. https://BioRender.com/x92914j . B Representative fluorescence images of macrophages at 0 day, 7 day, 15 day post-cisplatin exposure showing changes in the quantity and morphology distribution of macrophages (Hair cells: purple, macrophages: green). Typical macrophages were indicated with arrows respectively, stellate appearance in the apical (white), dendritic appearance in the middle (blue), and ameboid appearance at the basal (red), scale bar = 20 μm. C Calculations on basilar membrane confocal image stacks, using outer hair cells as the origin, with the direction toward the cochlear modiolus, respectively present the number of macrophages from the apical, middle, and basal turns of the cochlea within each 0.1 mm × 0.1 mm region after cisplatin treatment 7 day and 15 day ( n = 3 mice for each group). D Representative fluorescence images of macrophages and spiral ganglion neurons (SGNs) in frozen sections of CX3CR1 GFP/+ mice (SGN: green, CX3CR1: red), scale bar = 40 μm. E Representative fluorescence images of different macrophage marker labels (including CD68, Iba-1, CX3CR1, CD206, and CD163), scale bar = 40 μm. F Quantitative analysis of macrophage proportion expressing different markers within the cochlea on the 7-day post-cisplatin treatment ( n = 3 mice for each group). Statistical analyses were carried out via two-way ANOVA for ( C ) and t -test for ( F ). The data were presented as mean ± SD.* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: The primary antibodies used were as follows: F4/80 (CST, #30325S, 1:200), GFP polyclonal antibody (CST, #2955S, 1:500), anti-neurofilament heavy polypeptide (Abcam, #ab207176, 1:100), Myosin VIIa (a marker for hair cells, Proteus Biosciences, #25–6790, 1:500), anti-Iba-1 monoclonal antibody (Huabio, #ET1705-78, 1:200), DsRed (Santa Cruz, #sc-365989, 1:100), mcam (CST, #81701 T, 1:50), CD68 (Novus, #NB100-683, USA, 1:50), CD206 (Abcam, #ab64693, 1:200), CD163 (Abcam, #ab182422, 1:200), CD80 (Abcam, #ab315832, 1:100), anti-CtBP2 (BD, #612044, 1:200), anti-GluA2 (Merck Millipore, #MAB397, 1:200), and SF633 phalloidin (Solarbio, #CA1670-300T, 1:500).

Techniques: Fluorescence, Membrane, Marker, Expressing

Journal: Communications Biology

Article Title: Fortilin deficiency induces anti-atherosclerotic phenotypes in macrophages and protects hypercholesterolemic mice against atherosclerosis

doi: 10.1038/s42003-025-08425-w

Figure Lengend Snippet: A.U. arbitrary unit, RT-qPCR reverse transcription quantitative polymerase chain reaction, WT THP1 WT-fortilin cells, KO THP1 KO-fortilin cells, oxLDL oxidized low density lipoprotein, αSMA alpha smooth muscle cell actin, CD68 Cluster of Differentiation 68, TCE, 2,2,2-trichloroethanol total protein staining, GAPDH glyceraldehyde-3-phosphate dehydrogenase, phMΦ primary human macrophages, CTL control, phMΦ transduced by control short hairpin RNA (shRNA control ) lentiviral particles, KD knockdown, phMΦ transduced by shRNA fortilin lentiviral particles. a Expression levels of fortilin in WT and KO cells with and without oxLDL stimulation. b – f Status of expression of VSMC markers (αSMA, SM22α, CNN1) ( b – d ) and MФ markers (Mac2, CD68) ( e , f ) in WT and KO cells as analyzed by RT-qPCR (n = 3, 3). g Western blot analysis of VSMC and MΦ markers in WT and KO cells at their baselines and upon oxLDL stimulation. h Successful KD of fortilin using shRNA fortilin lentivirus particles in phMΦ. Status of expression of VSMC markers ( i – k ) and MΦ markers ( l , m ) in CTL and KD phMΦ cells as analyzed by RT-qPCR (n = 3, 3). Data are expressed as mean ± s.d., P values determined by one-way ANOVA with Tukey multiple comparisons ( a – f , h – m ) are shown. The findings in ( g ) were confirmed in three independent experiments.

Article Snippet: The following primary antibodies were used: Rabbit anti-fortilin (MBL International, Woburn, MA, USA; Catalog #: PM017; 1:1,000 dilution; used for the experiments depicted in Fig. ); Rabbit anti-fortilin (Abcam, Waltham, MA, USA; Clone EPR5540, ab133568; 1:2,000 dilution; Figs. , ); Mouse anti-GAPDH (Fitzgerald, Acton, MA, USA; Clone: 10R-G109a; 1:10,000 dilution; Fig. ); mouse anti-GAPDH (Santa Cruz Biotechnology, Inc. Dallas, TX, USA; Clone: 6C5; Catalog #: sc-32233; 1:1,000 dilution; Figs. , ); Rabbit anti-human calponin-1 (CNN1) mAb (Cell Signaling Technology (CST), Danvers, MA, USA; Clone D8L2T, Catalog #: 17819; 1:1,000 dilution; Fig. ); Recombinant rabbit anti-human α-smooth muscle actin (SMA) antibody (Abcam, clone EPR5368, Catalog #: ab124964; 1:1,000 dilution; Fig. ); Rabbit anti-human SM22α (Transgelin/TAGLN) polyclonal Ab (CST, Catalog #: 40471; 1:1,000 dilution; Fig. ); Rabbit anti-human TGF-β1 polyclonal Ab (Abcam, Catalog #: ab92486; 1:1,000 dilution; Figs. , ); Rabbit anti-Mac2 (Galectin-3/LGALS3) mAb (CST, Clone: D4I2R, Catalog #: 8785; 1:1,000 dilution; Fig. ); Rabbit anti-human CD68 mAb MultiMabTM(CST, multiple clones, Catalog #: 86985; 1:1,000 dilution; Fig. ).

Techniques: Quantitative RT-PCR, Reverse Transcription, Real-time Polymerase Chain Reaction, Staining, Control, shRNA, Knockdown, Expressing, Western Blot