Journal: Scientific Reports

Article Title: Species generalization and differences in Hedgehog pathway regulation of fungiform and circumvallate papilla taste function and somatosensation demonstrated with sonidegib

doi: 10.1038/s41598-018-34399-3

Figure Lengend Snippet: Sonidegib treatment reduces taste buds (TB), SHH ligand and proliferation in rat fungiform papilla (FP) while innervation is retained. ( a ) Immunofluorescent antibody detection of SHH ligand (red) and K19 (green) for TB cells; K18 (red) for TB cells and Ki67 (green) for cell proliferation, after Vehicle, 16d or 28d Sonidegib treatments. SHH is reduced in association with TB, K19+ cell loss. Asterisks (*) indicate nonspecific SHH immunoproduct in cornified surface cells in 16d Sonidegib image. The Vehicle, K18/Ki67 image shows 3 regions positive for Ki67+ cells (Apical, Basal and Perigemmal). Proliferating cells are lost in Apical FP region after 16–28d Sonidegib. ( b ) Number of Ki67+ cells in Apical and Basal regions of FP in Vehicle- and Sonidegib-treated mice. Numbers of tongues analyzed are in parentheses. For each tongue 8–10 FP were analyzed. Sonidegib treatment reduces apical epithelial cell proliferation in FP compared to Vehicle. Statistical analysis was one-way ANOVA with Tukey HSD posthoc comparisons (*p ≤ 0.05, compared to Vehicle, APICAL). ( c ) Immunofluorescent antibody detection of K19 or K18 (red) for TB cells and NF (green) for lingual and CT innervation or P2X3 (green) for CT nerve fibers. Innervation was retained after Sonidegib exposure. Asterisks (*) indicate nonspecific P2X3 immunoproduct in surface layer in Vehicle image. ( d ) Enlarged images from 28d Sonidegib papillae. Arrows point to NF+ or P2X3+ fibers in the FP epithelium. Throughout, white dotted lines indicate the basal lamina. Yellow dotted lines indicate surface of epithelium. ( a , c ) Scale bar: 50 μm, applies to all images. ( d ) Scale bar: 25 μm, applies to both images.

Article Snippet: Antibodies included: SHH ligand: goat anti-SHH (AF464, 0.1 μg/ml, R&D Systems for mice), mouse anti-SHH (sc-365112, 1:500, Santa Cruz Biotechnology for rat); taste cell markers K8, K18, K19 – , with different cytokeratins for optimal immunoreactions in double labeling, K8 (rat TROMA-1, 1:1000, Developmental Studies Hybridoma Bank) for mice, K18 (mouse anti-cytokeratin 18, sc-51582, 1:100, Santa Cruz Biotechnology) or K19 (rabbit anti-keratin 19, ab52625, 1:5000, Abcam) for rat; cell proliferation, Ki67 (rabbit anti-Ki67, NB110-89717SS, 1:5000, Novus Biologicals); innervation, NF (chicken anti-neurofilament heavy, ab4680, 1:5000, Abcam), P2X3 (rabbit anti-P2X3, APR-016, 1:1000, Alomone Labs).

Techniques:

Journal: Scientific Reports

Article Title: Species generalization and differences in Hedgehog pathway regulation of fungiform and circumvallate papilla taste function and somatosensation demonstrated with sonidegib

doi: 10.1038/s41598-018-34399-3

Figure Lengend Snippet: Sonidegib treatment in rat reduces taste buds (TB) and SHH ligand in circumvallate papilla (CV) whereas cell proliferation and GL innervation are retained. Immunofluorescent antibody detection of SHH ligand (red) and K19 (green) for TB cells; K18 (red) for TB cells and Ki67 (green) for cell proliferation; K19 (red) for TB cells and NF (green) for innervation; and, K18 (red) for TB cells and P2X3 (green) for taste nerve fibers, after Vehicle or 36d Sonidegib treatment. White dotted lines outline the epithelium. Asterisk in SHH/K19, 36d Sonidegib indicates nonspecific K19 immunostaining. Arrow points to the P2X3+ nerves extending into CV epithelium after Sonidegib treatment. SHH is reduced in association with TB cells. Cell proliferation is maintained and nerves fibers are retained after Sonidegib treatment. Scale bar: 50μm, applies to all images, except K19/NF.

Article Snippet: Antibodies included: SHH ligand: goat anti-SHH (AF464, 0.1 μg/ml, R&D Systems for mice), mouse anti-SHH (sc-365112, 1:500, Santa Cruz Biotechnology for rat); taste cell markers K8, K18, K19 – , with different cytokeratins for optimal immunoreactions in double labeling, K8 (rat TROMA-1, 1:1000, Developmental Studies Hybridoma Bank) for mice, K18 (mouse anti-cytokeratin 18, sc-51582, 1:100, Santa Cruz Biotechnology) or K19 (rabbit anti-keratin 19, ab52625, 1:5000, Abcam) for rat; cell proliferation, Ki67 (rabbit anti-Ki67, NB110-89717SS, 1:5000, Novus Biologicals); innervation, NF (chicken anti-neurofilament heavy, ab4680, 1:5000, Abcam), P2X3 (rabbit anti-P2X3, APR-016, 1:1000, Alomone Labs).

Techniques: Immunostaining

Journal: Scientific Reports

Article Title: Species generalization and differences in Hedgehog pathway regulation of fungiform and circumvallate papilla taste function and somatosensation demonstrated with sonidegib

doi: 10.1038/s41598-018-34399-3

Figure Lengend Snippet: Long term Sonidegib treatment in mouse reduces taste buds (TB) and SHH ligand in circumvallate papilla (CV) whereas proliferation and innervation are retained. Immunofluorescent antibody detection of SHH ligand (red) and K8 (green) for TB cells; K8 (red) for TB cells and Ki67 (green) for cell proliferation; and K8 (red) with NF (green) for GL innervation or P2X3 (green) for GL taste fibers, after Vehicle or 48d Sonidegib treatment. For SHH/K8, large dotted lines indicate the basal lamina. Small dotted lines outline the surface epithelium. Inset (K8/P2X3) shows an image of nerves extending into CV epithelial basal lamina (arrow). Scale bar: 50 μm, applies to all images. Inset at 2×.

Article Snippet: Antibodies included: SHH ligand: goat anti-SHH (AF464, 0.1 μg/ml, R&D Systems for mice), mouse anti-SHH (sc-365112, 1:500, Santa Cruz Biotechnology for rat); taste cell markers K8, K18, K19 – , with different cytokeratins for optimal immunoreactions in double labeling, K8 (rat TROMA-1, 1:1000, Developmental Studies Hybridoma Bank) for mice, K18 (mouse anti-cytokeratin 18, sc-51582, 1:100, Santa Cruz Biotechnology) or K19 (rabbit anti-keratin 19, ab52625, 1:5000, Abcam) for rat; cell proliferation, Ki67 (rabbit anti-Ki67, NB110-89717SS, 1:5000, Novus Biologicals); innervation, NF (chicken anti-neurofilament heavy, ab4680, 1:5000, Abcam), P2X3 (rabbit anti-P2X3, APR-016, 1:1000, Alomone Labs).

Techniques:

Journal: Neuro-Oncology Advances

Article Title: Double-modified oncolytic adenovirus armed with a recombinant interferon-like gene enhanced abscopal effects against malignant glioma

doi: 10.1093/noajnl/vdad117

Figure Lengend Snippet: Exploration of the antitumor mechanism of YSCH-01. (A) Flow cytometry was used to detect the apoptosis of LN-18 tumor cells after treatment. (B) Under 10 MOI of virus treatments, the comparison of PI proportion among Annexin V-FITC-positive LN-18 cells. (C) Flow cytometry was used to detect the apoptosis of LN-229 tumor cells after treatment. (D) Under 10 MOI of virus treatments, the comparison of PI proportion among Annexin V-FITC-positive LN-229 cells. (E) Western Blot to detect makers related to cell apoptosis. (F) The representative immuno-histochemical staining images of CD31, Ki67, CD45 in U-118 MG xenograft tumor sections 13 days after drug treatments. Scale bar, 100 μm. (G) The representative TUNEL immunofluorescent staining images of U-118 MG xenograft tumor sections 13 days after drug treatments. Scale bar, 1 mm. n.s., not significant, * P < .05, ** P < .01, *** P < . 001, **** P < . 0001, one-way ANOVA followed by Tukey’s multiple comparisons test.

Article Snippet: The primary antibodies used were а-Ki67 (#ab15580, abcam, Cambridge), а-CD45 (#ab208022, abcam), and а-CD31 (#ab182981, abcam).

Techniques: Flow Cytometry, Virus, Comparison, Western Blot, Staining, TUNEL Assay

Journal: Acta Pharmaceutica Sinica. B

Article Title: A triple combination strategy for nasopharyngeal carcinoma: Aptamer-guided liposomal chemotherapy, engineered NK cells, and Fc-enhanced PD-L1 antibody therapy

doi: 10.1016/j.apsb.2025.10.007

Figure Lengend Snippet: S3-Lip-DOX demonstrates enhanced tumor accumulation, potent in vivo efficacy, and a favorable safety profile. (A) Schematic of the experimental design for the in vivo tumor targeting and biodistribution study. (B) Representative whole-body fluorescence imaging of 5-8F tumor-bearing mice at indicated time points after intravenous injection of free DiR, Lip-DiR, or S3-Lip-DiR. (C) Ex vivo fluorescence images of major organs and tumors harvested 24 h post-injection. The tumor is positioned in the center of the organ array. (D) Quantification of the ex vivo fluorescence intensity in major organs and tumors from (C). (E) Schematic of the treatment schedule for the in vivo antitumor efficacy study. (F) Representative images of tumors excised from each treatment group on Day 22. (G) Tumor growth curves (left) and final tumor weights (right) of 5-8F xenograft-bearing mice after treatment. (H) Representative images of Ki67 (proliferation) and TUNEL (apoptosis) staining in tumor sections from each group. Scale bar = 100 μm. (I) Serum biochemical analysis for markers of liver function (ALT, ALB), kidney function (UREA, CREA), and cardiac injury (CK, LDH) from treated mice at the end of the study. Data in (D, G, I) are presented as mean ± SD ( n = 5 for D and G; n = 3 for I). Statistical significance was determined by one-way ANOVA. ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001; ns, not significant.

Article Snippet: Primary antibodies against human calreticulin (27298-1-AP), Bax (50599-2-Ig), and Bcl-2 (12789-1-AP), Ki67 (27309-1-AP) and PD-L1 (17952-1-AP) were purchased from Proteintech Group, Inc. (Wuhan, China), human cleaved caspase3 (#9661) and caspase3 (#14220) were purchased from Cell Signaling Technology (Beverly, MA, USA), and HRP-conjugated goat anti-rabbit IgG (ZB-2301) and goat anti-mouse IgG (ZB-2305) were obtained from Zhongshan Golden Bridge Biotechnology Co., Ltd. (Beijing, China).

Techniques: In Vivo, Fluorescence, Imaging, Injection, Ex Vivo, TUNEL Assay, Staining

Journal: Acta Pharmaceutica Sinica. B

Article Title: A triple combination strategy for nasopharyngeal carcinoma: Aptamer-guided liposomal chemotherapy, engineered NK cells, and Fc-enhanced PD-L1 antibody therapy

doi: 10.1016/j.apsb.2025.10.007

Figure Lengend Snippet: S3-P-NK cells exhibit superior tumor-homing, potent antitumor efficacy, and systemic safety in vivo . (A) Schematic of the experimental design for the in vivo biodistribution study. (B) Representative whole-body near-infrared fluorescence imaging of 5-8F tumor-bearing mice at indicated time points after intravenous injection of various DiR-labeled NK cell groups. (C) Ex vivo fluorescence images of major organs and tumors harvested 24 h post-injection. The tumor is positioned in the center of the organ array. (D) Quantification of the ex vivo fluorescence intensity in major organs and tumors from (C). (E) Schematic of the treatment schedule for the in vivo antitumor efficacy study. (F) Representative images of tumors excised from each treatment group at the study endpoint on Day 22. (G) Tumor growth curves (left) and final tumor weights (right) of 5-8F xenograft-bearing mice following the indicated treatments. (H) Representative images of Ki67 (proliferation) and TUNEL (apoptosis) staining in tumor sections from each group. Scale bar = 100 μm. (I) Serum biochemical analysis for markers of liver function (ALT, ALB), kidney function (UREA, CREA), and cardiac injury (CK, LDH) from treated mice at the end of the study. Data in (D, G, I) are presented as mean ± SD ( n = 5 for D, G; n = 3 for I). Statistical significance was determined by one-way ANOVA. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗∗ P < 0.0001; ns, not significant.

Article Snippet: Primary antibodies against human calreticulin (27298-1-AP), Bax (50599-2-Ig), and Bcl-2 (12789-1-AP), Ki67 (27309-1-AP) and PD-L1 (17952-1-AP) were purchased from Proteintech Group, Inc. (Wuhan, China), human cleaved caspase3 (#9661) and caspase3 (#14220) were purchased from Cell Signaling Technology (Beverly, MA, USA), and HRP-conjugated goat anti-rabbit IgG (ZB-2301) and goat anti-mouse IgG (ZB-2305) were obtained from Zhongshan Golden Bridge Biotechnology Co., Ltd. (Beijing, China).

Techniques: In Vivo, Fluorescence, Imaging, Injection, Labeling, Ex Vivo, TUNEL Assay, Staining

Journal: Acta Pharmaceutica Sinica. B

Article Title: A triple combination strategy for nasopharyngeal carcinoma: Aptamer-guided liposomal chemotherapy, engineered NK cells, and Fc-enhanced PD-L1 antibody therapy

doi: 10.1016/j.apsb.2025.10.007

Figure Lengend Snippet: Triple-combination therapy demonstrates potent synergistic antitumor efficacy and favorably remodels the tumor immune microenvironment in vivo . (A) Schematic of the in vivo triple-combination therapy schedule, detailing the doses, administration routes, and timing for S3-Lip-DOX, Atezolizumab/IgG1, and S3-P-NK cells. (B) Tumor growth curves monitored in vivo following different treatments. (C) Representative images of tumors excised from all treatment groups at the study endpoint on Day 22. (D) Final tumor weights quantified from each treatment group at the end of the study. (E) Representative flow cytometry plots showing the infiltration of activated macrophages (F4/80 + CD11b + CD86 + ) and mature dendritic cells (CD11c + MHCII + CD86 + ) within the tumor microenvironment of treated mice. (F) Quantification of the percentage of activated macrophages and mature dendritic cells from the flow cytometry analysis shown in (E). (G) Representative multiplex immunofluorescence images of tumor sections from key treatment groups. Panels show tumor cell proliferation (Ki67, red), apoptosis (TUNEL, green), macrophage polarization (F4/80, red; CD86, green; CD206, yellow), and dendritic cell maturation (CD11c, red; MHC II, yellow; CD86, green). Nuclei are counterstained with DAPI (blue). Scale bar = 100 μm. (H‒J) Quantitative analysis of positive signal area in multiplex immunofluorescence images (G, S10): Ki67 + area/TUNEL + area ratio (H); CD86 + area (M1 macrophages) and CD206 + area (M2 macrophages) (I); CD86 + area (dendritic cells, DCs) (J). Data in (B, D, F, H‒J) are presented as mean ± SD ( n = 5). Statistical significance was determined by one-way ANOVA (D, F, H‒J) or two-way ANOVA (B). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001.

Article Snippet: Primary antibodies against human calreticulin (27298-1-AP), Bax (50599-2-Ig), and Bcl-2 (12789-1-AP), Ki67 (27309-1-AP) and PD-L1 (17952-1-AP) were purchased from Proteintech Group, Inc. (Wuhan, China), human cleaved caspase3 (#9661) and caspase3 (#14220) were purchased from Cell Signaling Technology (Beverly, MA, USA), and HRP-conjugated goat anti-rabbit IgG (ZB-2301) and goat anti-mouse IgG (ZB-2305) were obtained from Zhongshan Golden Bridge Biotechnology Co., Ltd. (Beijing, China).

Techniques: In Vivo, Flow Cytometry, Multiplex Assay, Immunofluorescence, TUNEL Assay

Journal: Science signaling

Article Title: Multiple cancers escape from multiple MAPK pathway inhibitors and use DNA replication stress signaling to tolerate aberrant cell cycles.

doi: 10.1126/scisignal.ade8744

Figure Lengend Snippet: Fig. 6. Matrigel-embedded spheroids and clinical data highlight replication stress tolerance features of escapees. (A) Spheroid growth of the indicated cells after the indicated treatments. Across all spheroid panels: KRASi, 100 nM AMG510; BRAFi, 1 μM encorafenib; ATRi, 5 μM AZD6738; FANCi, 250 nM PIP199. Data are means ± SEM of 10 spheroids per condition, representative of two experiments; P values specified, determined by unpaired t test of the final time point. (B) Representative maximum intensity projections of 3D spheroid clusters stained for p-Rb and cleaved PARP (c-PARP, apoptotic marker) after 7 days of the indicated treatments. Scale bar, 20 μm. Images are representative of two experiments. (C) Quantification of cell count at day 7 in each individual spheroid. Each stacked bar represents one spheroid, where the cell count is broken down by p-Rb status. Combination treatment groups often resulted in spheroids with zero p-Rb+ cells. n = 5 spheroids per condition, representative of two experiments. (D) Percentage of c-PARP–positive cells per spheroid in (C). P values specified, determined by unpaired t test. (E) Formalin-fixed paraffin-embedded (FFPE) tissue sections longitudinally biopsied before and after treatment from two patients with BRAFV600E melanoma (see fig. S12A for patient and tumor details). Immunofluorescence of Ki67 (proliferation marker) and γ-H2AX (DNA damage marker) is displayed. Scale bars, 25 μm (MB2282) and 50 μm (MB3022). (F) Quantification of γ-H2AX intensity (blue coloring) in all cells and in Ki67+ cells (red coloring) from data in (E). Line drawn at the mean; n > 235 cells per ungated condition, n > 45 cells per Ki67-gated condition; P value as indicated or **P < 0.01 and ***P < 0.001, determined by permutation test. (G) Kaplan-Meier survival analyses based on data generated from the Harmonized Cancer Datasets within the Genomic Data Commons (GDC) research database (as of 28 April 2022; https://portal.gdc.cancer.gov). Regardless of treatment type, all patients with various tumor types harboring ≥1 EGFR, KRAS, or BRAF mutation and ≥1 TP53 mutation were included; N = 33 to 829 patients in each group. Cohort comparisons were made between tumor subgroups containing wild-type or mutant forms of the indicated genes. Leftmost plot examines a cohort that has ≥1 mutation among a list of FA-associated genes (fig. S13A). Displayed P values were determined by log-rank test.

Article Snippet: 16, eade8744 (2023) 1 August 2023 12 of 18 (D3U1W) mouse mAb (1:1000, Cell Signaling Technology, no. 86298); Ki67 sheep pAb (1:200, R&D Systems, no. AF7617); ERK1/2 mouse mAb (1:1000, Cell Signaling Technology, no. 4696); p-ERK1/2 (T202/Y204) mouse mAb (1:1000, MilliporeSigma, no. M9692); histone H3 mouse mAb (1:2000, Cell Signaling Technology, no. 3638); and cleaved PARP (Asp214) mouse mAb (1:500, BD Biosciences no. 552596).

Techniques: Staining, Marker, Cell Counting, Formalin-fixed Paraffin-Embedded, Immunofluorescence, Generated, Mutagenesis