Journal: Advanced Science

Article Title: A Novel FGFR3‐Targeting Antibody‐Drug Conjugate Induces Tumor Cell Apoptosis through the cGAS–STING Pathway in Bladder Cancer

doi: 10.1002/advs.202509933

Figure Lengend Snippet: A2 induces apoptosis in BC cells by targeting MAD2L1. A) Reaction scheme for biotinylation at the 20th hydroxyl group of A2. B) Overview of differentially expressed proteins identified by MS. C) Top 10 proteins ranked by the A2‐biotin/biotin abundance ratio. D) Western blot analysis of MAD2L1 expression in pull‐down products. E) Western blot detection of soluble MAD2L1 in T24 cells treated with A2 at different temperatures ( n = 3). F) SPR analysis of A2‐MAD2L1 binding kinetics and KD value. G) Representative immunofluorescence images showing co‐localization of A2‐biotin and MAD2L1 in T24 cells after 3 h of A2‐biotin treatment. Red: Anti‐MAD2L1, green: A2‐biotin, blue: DAPI, scale bar = 20 µm. H) Western blot analysis of MAD2L1 expression in T24 and UMUC‐3 cells treated with different concentrations of A2 for 24 h. I) Western blot validation of MAD2L1 OE in T24 and UMUC‐3 cells. J) Western blot validation of MAD2L1 KD in T24 and UMUC‐3 cells. K) Colony formation capacity of MAD2L1‐OE and control cells treated with different concentrations of A2 for 48 h. L) Apoptotic effects in MAD2L1‐OE and control cells treated with different concentrations of A2 for 48 h. ( n = 3). M) Apoptotic effects in MAD2L1‐KD and control cells treated with different concentrations of A2 for 48 h ( n = 3). Data are presented as mean ± SD. **** P < 0.0001. Statistical significance was determined by two‐way ANOVA followed by Tukey's multiple comparison test (L,M).

Article Snippet: SPR experiments were performed using a CM5 chip with amine coupling to immobilize MAD2L1 protein (HY‐ P71540 , MCE, USA).

Techniques: Western Blot, Expressing, Binding Assay, Immunofluorescence, Biomarker Discovery, Control, Comparison

Journal: Advanced Science

Article Title: A Novel FGFR3‐Targeting Antibody‐Drug Conjugate Induces Tumor Cell Apoptosis through the cGAS–STING Pathway in Bladder Cancer

doi: 10.1002/advs.202509933

Figure Lengend Snippet: A2 specifically binds to the Lys73 site of MAD2L1. A) Molecular docking simulation of the interaction between A2 and MAD2L1. B) Western blot validation of MAD2L1 expression levels in control, MAD2L1‐KD, and mutant T24 cells. C) Western blot analysis of MAD2L1 expression in pull‐down products from each group. D–F CETSA evaluating the binding of A2 to MAD2L1 in MAD2L1‐KD T24 cells reconstituted with MAD2L1 WT (D), MAD2L1V55A+I62A+V69A (E), and MAD2L1K73A (F) ( n = 3). Data are presented as mean ± SD.

Article Snippet: SPR experiments were performed using a CM5 chip with amine coupling to immobilize MAD2L1 protein (HY‐ P71540 , MCE, USA).

Techniques: Western Blot, Biomarker Discovery, Expressing, Control, Mutagenesis, Binding Assay

Journal: Advanced Science

Article Title: A Novel FGFR3‐Targeting Antibody‐Drug Conjugate Induces Tumor Cell Apoptosis through the cGAS–STING Pathway in Bladder Cancer

doi: 10.1002/advs.202509933

Figure Lengend Snippet: A2 targets MAD2L1 to activate the cGAS‐STING pathway. A) Effect of A2 on aneuploidy formation in T24 and UMUC‐3 cells. B) Representative images showing cGAS subcellular distribution and micronuclei formation (indicated by arrows) in T24 and UMUC‐3 cells treated with 0.064 µ m A2 for 48 h. Red: cGAS, green: PicoGreen (DNA stain), Scale bar = 20 µm. C) Quantification of cytosolic DNA in T24 and UMUC‐3 cells treated with 0.32 µ m A2 for 48 h ( n = 3). D) Western blot analysis of cGAS‐STING pathway protein expression in T24 and UMUC‐3 cells treated with different concentrations of A2 for 48 h. E) Western blot analysis of cGAS‐STING pathway protein expression in MAD2L1‐OE and control cells treated with different concentrations of A2 for 48 h. F) Western blot analysis of cGAS‐STING pathway protein expression in MAD2L1‐KD and control cells treated with different concentrations of A2 for 48 h. G) Apoptotic effects in T24 and UMUC‐3 cells treated with 1.6 µ m A2 after STING inhibitor H151‐mediated blockade of the cGAS‐STING pathway. Data are presented as mean ± SD. **** P < 0.0001. Statistical significance was determined by a two‐tailed Student's t‐ test (C).

Article Snippet: SPR experiments were performed using a CM5 chip with amine coupling to immobilize MAD2L1 protein (HY‐ P71540 , MCE, USA).

Techniques: Staining, Western Blot, Expressing, Control, Two Tailed Test

Journal: Advanced Science

Article Title: A Novel FGFR3‐Targeting Antibody‐Drug Conjugate Induces Tumor Cell Apoptosis through the cGAS–STING Pathway in Bladder Cancer

doi: 10.1002/advs.202509933

Figure Lengend Snippet: Cytotoxicity of LZU‐WZLYCS01 against BC cells and PDOs, and validation of the bystander effect. A) Cell viability of WT and FGFR3‐KO T24 and UMUC‐3 cells treated with different concentrations of LZU‐WZLYCS01, A2, or FGFR3 antibody for 72 h. B) Apoptotic effects in WT and FGFR3‐KO T24 and UMUC‐3 cells treated with different concentrations of LZU‐WZLYCS01 for 48 h ( n = 3). C) Colony formation capacity of T24 and UMUC‐3 cells treated with LZU‐WZLYCS01 for 48 h. D) Schematic diagram of the co‐culture model for assessing the bystander effects. E) Effect of LZU‐WZLYCS01 on the viability of FGFR3‐KO T24 and UMUC‐3 cells in the co‐culture system ( n = 3). F) Representative bright‐field and AM/PI‐stained images showing LZU‐WZLYCS01‐induced cytotoxicity in PDOs. Green: viable cells, red: dead cells, scale bar = 50 µm. G) Representative H&E staining of PDOs and matched parental tumor tissues. Scale bar = 25 µm. H) Representative IHC staining of UPK2, FGFR3, and MAD2L1 in PDOs. Scale bar = 25 µm. Data are presented as mean ± SD. * P < 0.05, ** P < 0.01, **** P < 0.0001. Statistical significance was determined by two‐way ANOVA followed by Tukey's multiple comparison test (B) and a two‐tailed Student's t‐ test (E).

Article Snippet: SPR experiments were performed using a CM5 chip with amine coupling to immobilize MAD2L1 protein (HY‐ P71540 , MCE, USA).

Techniques: Biomarker Discovery, Co-Culture Assay, Staining, Immunohistochemistry, Comparison, Two Tailed Test

Journal: Advanced Science

Article Title: A Novel FGFR3‐Targeting Antibody‐Drug Conjugate Induces Tumor Cell Apoptosis through the cGAS–STING Pathway in Bladder Cancer

doi: 10.1002/advs.202509933

Figure Lengend Snippet: Mechanism of action and tumor‐targeting capability of LZU‐WZLYCS01. A) Flow cytometry analysis of LZU‐WZLYCS01 binding and internalization in T24 and UMUC‐3 cells at 37 and 4 °C. B) Representative images showing LZU‐WZLYCS01 binding, internalization, and lysosomal co‐localization in T24 and UMUC‐3 cells at 37 and 4 °C. Red: LZU‐WZLYCS01, green: anti‐LAMP2, blue: DAPI, scale bar = 10 µm. C) Western blot analysis of cGAS‐STING pathway protein expression in T24 and UMUC‐3 cells treated with different concentrations of LZU‐WZLYCS01 for 48 h. D) Western blot analysis of cGAS‐STING pathway protein expression in MAD2L1‐OE and control cells treated with different concentrations of LZU‐WZLYCS01 for 48 h. E) In vivo fluorescence imaging of UMUC‐3 xenograft models 24 h after intravenous injection of LZU‐WZLYCS01–Cy5 ( n = 3). F) Ex vivo fluorescence imaging of tumors and major organs (heart, liver, spleen, lungs, kidneys, and brain) collected 24 h post‐injection.

Article Snippet: SPR experiments were performed using a CM5 chip with amine coupling to immobilize MAD2L1 protein (HY‐ P71540 , MCE, USA).

Techniques: Flow Cytometry, Binding Assay, Western Blot, Expressing, Control, In Vivo, Fluorescence, Imaging, Injection, Ex Vivo

Journal: The Journal of Cell Biology

Article Title: Aurora A kinase phosphorylates Hec1 to regulate metaphase kinetochore–microtubule dynamics

doi: 10.1083/jcb.201707160

Figure Lengend Snippet: Hec1 S69 is phosphorylated throughout mitosis. (A) Amino acid sequences of the human and PtK1 cell Hec1 N-terminal tail domain. Shown in yellow is the human peptide sequence that was used to generate the S69 phosphospecific antibody. The arrow points to S69 in the human sequence and the corresponding serine residue in the PtK1 sequence. Asterisks indicate all other mapped Aurora B kinase sites in the human Hec1 tail domain ( ; ). (B) Immunofluorescence images of HeLa cells stained with phosphospecific antibodies to Hec1 pS69. Depletion of Hec1 (bottom) results in loss of pS69 staining at kinetochores. Cells are also immunostained with antibody 9G3 (pan-Hec1 antibody) and an anticentromere antibody (ACA) derived from human calcinosis, Raynaud's phenomenon, esophageal dysmotility, sclerodactyly, and telangiectasia (CREST) patient serum. (C) Immunofluorescence images of HeLa cells demonstrating kinetochore localization of pS69 during mitosis. Quantification is shown on the right. For each phase shown, ≥400 kinetochores from ≥30 cells were measured. (D) Immunofluorescence images of a HeLa cell stained with antibodies to pS69 and Mad2. For the cell shown, most chromosomes are aligned at the spindle equator, and one chromosome remains near a spindle pole (arrows). A schematic illustrating examples of pole-proximal chromosomes is shown on the left. (E) Immunofluorescence images of HeLa cells depleted of CENP-E to increase the number of pole-proximal chromosomes (arrows) and stained with Hec1 phosphospecific antibodies. Quantification is shown on the right from one representative experiment. n values are as follows: pS69, 20 polar kinetochores and 40 aligned kinetochores; pS55, 17 polar kinetochores and 57 aligned kinetochores; and pS44, 13 polar kinetochores and 29 aligned kinetochores. Error bars indicate SD. Bars: (B, C, and E) 10 µm; (D) 3 µm.

Article Snippet: Two rabbits were immunized with the Mad2 protein (Rockland Immunochemicals Inc), and antisera from the two injected rabbits were affinity purified against full-length Mad2 protein using a HiTrap-NHS column.

Techniques: Sequencing, Residue, Immunofluorescence, Staining, Derivative Assay

Journal: BMC oral health

Article Title: 5'tiRNA-35-GlyTCC-3 and 5'tiRNA-33-CysGCA-11 target BMP6, CUL1 and SPR of non-syndromic cleft palate.

doi: 10.1186/s12903-025-05661-8

Figure Lengend Snippet: Fig. 6 Target three key gene loci of two key tsRNAs in NSCPO tissues. (a) 5’tiRNA-35-GlyTCC-3 targets BMP6. (b) 5’tiRNA-35-GlyTCC-3 targets CUL1. (c) 5’tiRNA-33-CysGCA-11 targets SPR

Article Snippet: Primary antibodies were employed in opposition to the subsequent proteins: BMP6 (M06924-1, Boster, China), SPR (bs-11784R, Bioss Antibodies, China) and CUL1 (K003246P, Solarbio, China).

Techniques:

Journal: BMC oral health

Article Title: 5'tiRNA-35-GlyTCC-3 and 5'tiRNA-33-CysGCA-11 target BMP6, CUL1 and SPR of non-syndromic cleft palate.

doi: 10.1186/s12903-025-05661-8

Figure Lengend Snippet: Fig. 9 Relative expression of three key mRNAs SPR (a), BMP6 (b) and CUL1 (c) in clinical samples by RT-qPCR. (**** p < 0.0001; ***p < 0.001; **p < 0.01)

Article Snippet: Primary antibodies were employed in opposition to the subsequent proteins: BMP6 (M06924-1, Boster, China), SPR (bs-11784R, Bioss Antibodies, China) and CUL1 (K003246P, Solarbio, China).

Techniques: Expressing, Quantitative RT-PCR

Journal: BMC oral health

Article Title: 5'tiRNA-35-GlyTCC-3 and 5'tiRNA-33-CysGCA-11 target BMP6, CUL1 and SPR of non-syndromic cleft palate.

doi: 10.1186/s12903-025-05661-8

Figure Lengend Snippet: Fig. 10 Immunohistochemistry staining of BMP6, CUL1, SPR in the Health, Ctrl and NSCPO tissue

Article Snippet: Primary antibodies were employed in opposition to the subsequent proteins: BMP6 (M06924-1, Boster, China), SPR (bs-11784R, Bioss Antibodies, China) and CUL1 (K003246P, Solarbio, China).

Techniques: Immunohistochemistry, Staining