Journal: Cancers

Article Title: Optimizing Sequential Targeted Therapies in Advanced Renal Cell Carcinoma Using Patient-Derived Orthotopic Xenograft Mouse Avatars

doi: 10.3390/cancers18101615

Figure Lengend Snippet: Histopathological comparison of tumor architecture and proliferation between patient renal cell carcinoma (RCC) specimens and corresponding patient-derived xenograft models. Formalin-fixed, paraffin-embedded sections from primary patient tumors (KiCa-Pt58 and KiCa-Pt118), matched subcutaneous xenografts, and intra-renal patient-derived orthotopic xenograft (PDOX) tumors were analyzed to assess preservation of tumor histology and proliferative characteristics. ( A , C ) Representative images of KiCa-Pt58 ( A ) and KiCa-Pt118 ( C ) tissues stained with hematoxylin and eosin (H&E) for tissue architecture or were subjected to immunohistochemistry (IHC) for human CD44 (tumor cell marker) and human Ki67 (proliferation marker). Brown staining indicates positive immunoreactivity. Images were captured at 100× original magnification using an Axiovert 200M deconvolution microscope and SlideBook 6.0 software (Intelligent Imaging Innovations, Denver, CO, USA). ( B , D ) Quantitative analysis of Ki67-positive area (%) in KiCa-Pt58 ( B ) and KiCa-Pt118 ( D ) tissues. Positive (brown) staining areas were quantified digitally using Adobe Photoshop 7.0, with the percentage of immunoreactive area calculated per field. Data are presented as mean ± SEM (multiple fields per sample). Comparisons among patient biopsy specimens, subcutaneous xenografts, and orthotopic PDOX tumors were performed using unpaired Student’s t -test. No significant differences were observed (ns, p > 0.05), demonstrating faithful recapitulation of the parental tumor proliferative index across model passages. Abbreviations: RCC, renal cell carcinoma; PDOX, patient-derived orthotopic xenograft; H&E, hematoxylin and eosin; IHC, immunohistochemistry.

Article Snippet: Paraffin-embedded sections (5 μm) were stained with H&E or subjected to immunohistochemistry using primary antibodies against human Ki67 (proliferation marker; Thermo Fisher Scientific, Waltham, MA, USA; 1:200), human CD44 (tumor cell marker; Acris Antibodies, Rockville, MD, USA; 1:75), mouse CD31 (angiogenesis marker; Abcam, Cambridge, MA, USA; 1:200), and human PD-L1 (programmed death-ligand 1, immune checkpoint ligand; BioLegend, San Diego, CA, USA; 1:200) [ ].

Techniques: Comparison, Derivative Assay, Formalin-fixed Paraffin-Embedded, Preserving, Staining, Immunohistochemistry, Marker, Microscopy, Software, Imaging

Journal: Cancers

Article Title: Optimizing Sequential Targeted Therapies in Advanced Renal Cell Carcinoma Using Patient-Derived Orthotopic Xenograft Mouse Avatars

doi: 10.3390/cancers18101615

Figure Lengend Snippet: Histopathological and immunohistochemical evaluation of targeted therapy responses in the KiCa-Pt58 patient-derived orthotopic xenograft (PDOX) model. ( A ) Representative images of left kidney tumors from KiCa-Pt58 PDOX mice after vehicle control or sequential targeted therapy (Everolimus→Sunitinib [E→S], Pazopanib→Sunitinib [P→S], Sunitinib→Everolimus [S→E], Pazopanib→Everolimus [P→E]). Formalin-fixed, paraffin-embedded sections were stained with hematoxylin and eosin (H&E) for tumor architecture or subjected to immunohistochemistry (IHC) for human CD44 (tumor cell marker), human Ki67 (proliferation marker), mouse CD31 (angiogenesis/endothelial marker), and human PD-L1 (immune checkpoint ligand). Brown staining indicates positive immunoreactivity. Images were acquired at 100× original magnification using an Axiovert 200M deconvolution microscope and SlideBook 6.0 software (Intelligent Imaging Innovations, Denver, CO, USA). ( B – D ) Quantitative analysis of positive staining area (%) for Ki67 ( B ), CD31 ( C ), and PD-L1 ( D ) across treatment groups. Positive (brown) areas were quantified digitally using Adobe Photoshop 7.0 (percentage immunoreactive area per field). Data are presented as mean ± SEM (multiple fields per sample; n = 7–9 mice per group). Statistical comparisons vs. control were performed using one-way ANOVA followed by Dunnett’s or Tukey’s post hoc tests (GraphPad Prism v7). Asterisks indicate significance: * p < 0.05; ** p < 0.01; *** p < 0.001. Effective regimens (particularly P→E and S→E) significantly reduced Ki67+ proliferation, CD31+ vascularity, and PD-L1 expression compared to control, consistent with antitumor and potential immunomodulatory effects. Abbreviations: PDOX, patient-derived orthotopic xenograft; IHC, immunohistochemistry; E, everolimus; S, sunitinib; P, pazopanib.

Article Snippet: Paraffin-embedded sections (5 μm) were stained with H&E or subjected to immunohistochemistry using primary antibodies against human Ki67 (proliferation marker; Thermo Fisher Scientific, Waltham, MA, USA; 1:200), human CD44 (tumor cell marker; Acris Antibodies, Rockville, MD, USA; 1:75), mouse CD31 (angiogenesis marker; Abcam, Cambridge, MA, USA; 1:200), and human PD-L1 (programmed death-ligand 1, immune checkpoint ligand; BioLegend, San Diego, CA, USA; 1:200) [ ].

Techniques: Immunohistochemical staining, Derivative Assay, Control, Formalin-fixed Paraffin-Embedded, Staining, Immunohistochemistry, Marker, Microscopy, Software, Imaging, Expressing

Journal: Cancers

Article Title: Optimizing Sequential Targeted Therapies in Advanced Renal Cell Carcinoma Using Patient-Derived Orthotopic Xenograft Mouse Avatars

doi: 10.3390/cancers18101615

Figure Lengend Snippet: Histopathological and immunohistochemical assessment of targeted therapy responses in the KiCa-Pt118 patient-derived orthotopic xenograft (PDOX) model. ( A ) Representative images of left kidney tumors from KiCa-Pt118 PDOX mice after vehicle control or sequential targeted therapy (Everolimus→Sunitinib [E→S], Pazopanib→Sunitinib [P→S], Sunitinib→Everolimus [S→E], Pazopanib→Everolimus [P→E]). Formalin-fixed, paraffin-embedded sections were stained with hematoxylin and eosin (H&E) for tumor architecture or subjected to immunohistochemistry (IHC) for human CD44 (tumor cell marker), human Ki67 (proliferation marker), mouse CD31 (angiogenesis/endothelial marker), and human PD-L1 (immune checkpoint ligand). Brown staining indicates positive immunoreactivity. Images were acquired at 100× original magnification using an Axiovert 200M deconvolution microscope and SlideBook 6.0 software (Intelligent Imaging Innovations, Denver, CO, USA). ( B – D ) Quantitative analysis of positive staining area (%) for Ki67 ( B ), CD31 ( C ), and PD-L1 ( D ) across treatment groups. Positive (brown) areas were quantified digitally using Adobe Photoshop 7.0 (percentage immunoreactive area per field), as described in . Data are presented as mean ± SEM (multiple fields per sample; n = 7–9 mice per group). Statistical comparisons vs. control were performed using one-way ANOVA followed by Dunnett’s or Tukey’s post hoc tests (GraphPad Prism v7). Asterisks indicate significance: * p < 0.05; ** p < 0.01; *** p < 0.001. Effective regimens (particularly S→E) significantly reduced Ki67+ proliferation, CD31+ vascularity, and PD-L1 expression compared to control, consistent with antitumor activity in this indolent, non-metastatic model. Abbreviations: PDOX, patient-derived orthotopic xenograft; IHC, immunohistochemistry; E, everolimus; S, sunitinib; P, pazopanib.

Article Snippet: Paraffin-embedded sections (5 μm) were stained with H&E or subjected to immunohistochemistry using primary antibodies against human Ki67 (proliferation marker; Thermo Fisher Scientific, Waltham, MA, USA; 1:200), human CD44 (tumor cell marker; Acris Antibodies, Rockville, MD, USA; 1:75), mouse CD31 (angiogenesis marker; Abcam, Cambridge, MA, USA; 1:200), and human PD-L1 (programmed death-ligand 1, immune checkpoint ligand; BioLegend, San Diego, CA, USA; 1:200) [ ].

Techniques: Immunohistochemical staining, Derivative Assay, Control, Formalin-fixed Paraffin-Embedded, Staining, Immunohistochemistry, Marker, Microscopy, Software, Imaging, Expressing, Activity Assay

Journal: bioRxiv

Article Title: Desmoglein-3 modulates p38MAPK and ERK signaling responses through the mechano-sensitive channel Piezo1

doi: 10.64898/2026.05.11.723746

Figure Lengend Snippet: A . Immunofluorescence staining for DSG3 and desmoplakin (DSP) of CTRL and DSG3-KO HaCaT keratinocytes. Scale bar 10 μm. B . Quantification of DSP intensity over the corresponding membrane length (μm), Mann-Whitney test. Each data point represents one cell from three independent experiments in total. C . Immunofluorescence staining for Pan-cytokeratin (pan-CK) of CTRL and DSG3-KO HaCaT keratinocytes. Scale bar 10 μm. D . Analysis of pan-CK staining by quantifying the intensity (a.u) perpendicular to cell borders over a distance of 10 µm. E . Dispase-based dissociation assay of CTRL and DSG3-KO cells. Representative images and quantifications of N = 3 are shown, Welch’s t-test. F-G . Western blot and representative quantification of phosphorylated p38MAPK (p-p38MAPK) and ERK (pERK) in CTRL and DSG3-KO HaCaT keratinocytes. Each dot represents biological replicates. Welch’s t-test.

Article Snippet: The following primary antibodies were diluted with Odyssey blocking buffer in TBS containing 0.1% Tween 20 (Thermo Fisher Scientific) and incubated overnight at 4°C, with rotation: mouse GAPDH mAb (Santa Cruz Biotechnology, #sc-47724), rabbit phospho-p38MAPK Thr180/Tyr182 (Cell Signaling Technology, #4511S), rabbit p38MAPK (Cell Signaling Technology, #9212S), rabbit ERK 1/2 (p44/42) (Cell Signaling Technology, #9102), phospho-ERK (Santa Cruz, sc-7383), mouse plakoglobin (Progen, #61005), mouse desmoplakin 1/2 (Progen, #61003), rabbit Piezo1 (Proteintech, # 15939-1-AP), rabbit Desmoglein-3 (5G11, Santa Cruz Biotechnology, # sc-53487), Desmoglein-2 (10G11, Acris OriGene, #BM5016), mouse Pan Cytokeratin mAb (Thermo Fisher, #41-9003-82), Phalloidin (Thermo Fisher, #21833).

Techniques: Immunofluorescence, Staining, Membrane, MANN-WHITNEY, Western Blot

Journal: The Journal of Pathology: Clinical Research

Article Title: The prognostic significance of the IASLC grading system in ALK ‐positive invasive non‐mucinous adenocarcinoma of the lung: correlation with clinicopathological features

doi: 10.1002/2056-4538.70093

Figure Lengend Snippet: Increased IASLC grade association with elevated immune cell infiltration. (A) H&E staining showing increased TILs with grade elevation. (B) Higher IASLC grades exhibited a rise in intratumoral CD4+ T‐cell accumulation. (C) Correlation of advanced IASLC grades with a prominent rise in CD8+ T‐lymphocytes. (D) Lack of significant difference in FoxP3 + lymphocyte count across grades. (E) Quantitative analysis of CD4+, CD8+, and FoxP3+ cells by IHC. (F) CD4:CD8 and CD8:FoxP3 ratios across IASLC grades (bar = 50 μm). H&E, hematoxylin and eosin; IASLC, International Association for the Study of Lung Cancer; TILs, tumor‐infiltrating lymphocytes.

Article Snippet: The antibodies used included: ALK (790‐4797, ready‐to‐use, Ventana®), TTF‐1 (11833669A, 1:100, Zhongshan Golden Bridge Biotechnology, Beijing, China), p40 (25081519, 1:100, Dako, Carpinteria, CA, USA), Ki‐67 ( GT209429 , 1:100, Gene Tech Company Limited, Shanghai, China), PD‐L1 (SK006, ready‐to‐use, Dako, Carpinteria, CA, USA), CD4 (IR649, 1:100, Dako, Carpinteria, CA, USA), CD8 (ZA‐0508, 1:100, Zhongshan Golden Bridge Biotechnology, Beijing, China), and FoxP3 (sc‐53876, 1:100, Santa Cruz, Dallas, TX, USA).

Techniques: Staining