Journal: Regenerative Therapy

Article Title: WTAP stabilizes MMP12 expression to promote the malignant phenotypes of esophageal cancer cells

doi: 10.1016/j.reth.2026.101101

Figure Lengend Snippet: MMP12 silencing inhibited M2 macrophage polarization. THP-1 cells were differentiated into M0 macrophages (THP-1 M0) by treatment with 100 ng/mL PMA for 24 h. (A) Flow cytometry was used to quantify the number of CD68-positive cells. Subsequently, KYSE150 cells were co-cultured with the THP-1-derived macrophages using a Transwell system. (B) The mRNA levels of IL-10, Arg-1, and TGF-β were detected by qRT-PCR. (C) Flow cytometry was used to quantify the number of CD206-positive macrophages. (D) Cell migration analysis by transwell migration assay. ∗ P < 0.05, ∗∗ P < 0.01 and ∗∗∗ P < 0.001.

Article Snippet: For immunophenotyping, single-cell suspensions of both THP-1 and THP-1-M0 cells were labeled with an anti-CD68 antibody (E-AB-F1299L, Elabscience, Wuhan, China).

Techniques: Flow Cytometry, Cell Culture, Derivative Assay, Quantitative RT-PCR, Migration, Transwell Migration Assay

Journal: Regenerative Therapy

Article Title: WTAP stabilizes MMP12 expression to promote the malignant phenotypes of esophageal cancer cells

doi: 10.1016/j.reth.2026.101101

Figure Lengend Snippet: WTAP silencing inhibited M2 macrophage polarization by regulating MMP12. THP-1 cells were differentiated into M0 macrophages (THP-1 M0) by treatment with 100 ng/mL PMA for 24 h. KYSE150 cells were transfected with si-WTAP, MMP12 overexpression plasmid, or the matched control (si-NC and oe-NC). Subsequently, these KYSE150 cells were co-cultured with the THP-1-derived macrophages using a Transwell system. (A) The mRNA levels of IL-10, Arg-1, and TGF-β were detected by qRT-PCR. (B) Flow cytometry was used to quantify the number of CD206-positive macrophages. (C) Cell migration analysis by transwell migration assay. ∗ P < 0.05, ∗∗ P < 0.01 and ∗∗∗ P < 0.001.

Article Snippet: For immunophenotyping, single-cell suspensions of both THP-1 and THP-1-M0 cells were labeled with an anti-CD68 antibody (E-AB-F1299L, Elabscience, Wuhan, China).

Techniques: Transfection, Over Expression, Plasmid Preparation, Control, Cell Culture, Derivative Assay, Quantitative RT-PCR, Flow Cytometry, Migration, Transwell Migration Assay

Journal: Redox Biology

Article Title: Peroxiredoxin Ⅲ safeguards cardiac function against doxorubicin by regulating mitochondrial quality control via H 2 O 2 detoxification

doi: 10.1016/j.redox.2026.104176

Figure Lengend Snippet: Doxorubicin increases mitochondrial H 2 O 2 levels and causes cell death in rat cardiomyocytes. (A) H9c2 cells were treated with the indicated concentrations of DOX for 24 h. Cell death was assessed by Annexin V-FITC staining and analyzed using flow cytometer. Representative histograms of three independent experiments are shown. (B) H9c2 cells were treated with 1 μM DOX for the indicated times. Cell viability was measured using WST-1 reagent. (C-D) H9c2 cells were treated with 1 μM DOX for the indicated times and then stained with 5 μM CM-H 2 DCFDA (C) or 5 μM MitoPY-1 (D) for 15 min. Relative fluorescence intensity (RFI) was measured by flow cytometer. All data are presented as mean ± S.D. (n = 3). Statistical significance was determined by one-way ANOVA followed by Dunnett's post hoc test. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 compared to untreated control.

Article Snippet: Rat cardiomyocyte cell line, H9c2 cells (CRL-1446) were obtained from the American Type Culture Collection (Manassas, VA, USA) and cultured in DMEM high glucose (SH30022.01, Hyclone, Logan, UT, USA) supplemented with 10% FBS, 1% antibiotic-antimycotic, and 1 μg/ml puromycin.

Techniques: Staining, Flow Cytometry, Fluorescence, Control

Journal: Redox Biology

Article Title: Peroxiredoxin Ⅲ safeguards cardiac function against doxorubicin by regulating mitochondrial quality control via H 2 O 2 detoxification

doi: 10.1016/j.redox.2026.104176

Figure Lengend Snippet: PrxⅢ regulates mitochondrial H 2 O 2 accumulation and oxidative damage in DOX-treated cardiomyocytes. (A) H9c2 cells stably expressing either control (pSUPER) or PrxⅢ-targeting siRNA (pSUPER-siPrxⅢ) were transduced with Ad-PrxⅢ or Ad-Stuffer for 24 h and then exposed to 1 μM DOX for 9 h. Mitochondrial H 2 O 2 levels were assessed using the mitochondria-targeted fluorescent probe MitoPY-1 and visualized by green fluorescence. Scale bar, 75 μm. Five randomly selected microscopic fields per sample were analyzed and combined to represent one biological replicate. (B) Cells were transduced with Ad-PrxⅢ or Ad-Stuffer for 24 h and then exposed to 1 μM DOX for 18 h. FOXO3a localization was assessed by immunofluorescence using anti-FOXO3a (green) antibody and DAPI (blue). Scale bar, 25 μm Five randomly selected microscopic fields per sample were analyzed and combined to represent one biological replicate. All data are expressed as mean ± S.D. Statistical significance was determined using two-way ANOVA followed by Bonferroni's post hoc test. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 and ns, p > 0.05.

Article Snippet: Rat cardiomyocyte cell line, H9c2 cells (CRL-1446) were obtained from the American Type Culture Collection (Manassas, VA, USA) and cultured in DMEM high glucose (SH30022.01, Hyclone, Logan, UT, USA) supplemented with 10% FBS, 1% antibiotic-antimycotic, and 1 μg/ml puromycin.

Techniques: Stable Transfection, Expressing, Control, Transduction, Fluorescence, Immunofluorescence

Journal: Redox Biology

Article Title: Peroxiredoxin Ⅲ safeguards cardiac function against doxorubicin by regulating mitochondrial quality control via H 2 O 2 detoxification

doi: 10.1016/j.redox.2026.104176

Figure Lengend Snippet: PrxⅢ regulates oxidative damage and mitochondrial bioenergetics in DOX-treated cardiomyocytes. (A) Cells were transduced with Ad-PrxⅢ or Ad-Stuffer for 24 h and then exposed to 1 μM DOX for 12 h. Cells were labeled with 5 μM 10-NAO and relative fluorescence intensity (RFI) was analyzed by flow cytometer. Representative histograms and quantification are shown. (B) Cells were treated with 1 μM DOX for 12 h and labeled with 10 μM Rho-123 to assess mitochondrial membrane potential ( ΔΨ m ). The percentage of cells with low ΔΨ m was quantified by flow cytometry. Representative histograms and quantification are shown. (C) Cells were transduced with Ad-PrxⅢ or Ad-Stuffer for 24 h and then exposed to 1 μM DOX for 12 h. Representative bar graph showing relative ATP concentration normalized to protein contents. (D-H) Cells were transduced with Ad-PrxⅢ or Ad-Stuffer for 24 h and then exposed to 1 μM DOX for 12 h. (D) Representative tracing of the oxygen consumption rate (OCR). Arrows indicate time points when cells were treated with oligomycin, FCCP, and rotenone plus antimycin A (Rot/AA), respectively. The OCR is calculated as (E) basal respiration, (F) maximal respiration, (G) spare respiration, and (H) proton-leak. All data are expressed as mean ± S.D. (n = 3-5). Statistical significance was determined using two-way ANOVA followed by Bonferroni's post hoc test. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001.

Article Snippet: Rat cardiomyocyte cell line, H9c2 cells (CRL-1446) were obtained from the American Type Culture Collection (Manassas, VA, USA) and cultured in DMEM high glucose (SH30022.01, Hyclone, Logan, UT, USA) supplemented with 10% FBS, 1% antibiotic-antimycotic, and 1 μg/ml puromycin.

Techniques: Transduction, Labeling, Fluorescence, Flow Cytometry, Membrane, Concentration Assay

Journal: Redox Biology

Article Title: Peroxiredoxin Ⅲ safeguards cardiac function against doxorubicin by regulating mitochondrial quality control via H 2 O 2 detoxification

doi: 10.1016/j.redox.2026.104176

Figure Lengend Snippet: PrxⅢ regulates mitochondrial biogenesis in DOX-treated cardiomyocytes. (A) Cells were transduced with Ad-PrxⅢ or Ad-Stuffer for 24 h and then exposed to 1 μM DOX for 6 h. Total RNA was isolated and the mRNA levels of PGC-1α, Nrf1, and mtTFA were determined by qRT-PCR. Representative bar graphs showing relative expression normalized to GAPDH are presented. (B) Relative mRNA expression of the mitochondrial DNA-encoded cytochrome c oxidase (COX) mRNA was measured as an indicator of mtDNA copy-dependent transcription. Representative bar graphs showing relative expression normalized to GAPDH are presented. (C) Cells were transduced with Ad-PrxⅢ or Ad-Stuffer for 24 h and then exposed to 1 μM DOX for 18 h. Representative immunoblots and quantitative analyses of PGC-1α, Nrf1, and mtTFA proteins. GAPDH and α-tubulin were used as loading controls. Densitometric quantification of protein expression was performed from three independent biological replicates. All data are presented as mean ± S.D. (n = 3). Statistical significance was determined using two-way ANOVA followed by Bonferroni's post hoc test. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001.

Article Snippet: Rat cardiomyocyte cell line, H9c2 cells (CRL-1446) were obtained from the American Type Culture Collection (Manassas, VA, USA) and cultured in DMEM high glucose (SH30022.01, Hyclone, Logan, UT, USA) supplemented with 10% FBS, 1% antibiotic-antimycotic, and 1 μg/ml puromycin.

Techniques: Transduction, Isolation, Quantitative RT-PCR, Expressing, Western Blot

Journal: Redox Biology

Article Title: Peroxiredoxin Ⅲ safeguards cardiac function against doxorubicin by regulating mitochondrial quality control via H 2 O 2 detoxification

doi: 10.1016/j.redox.2026.104176

Figure Lengend Snippet: PrxⅢ protects cardiomyocytes against DOX-induced apoptotic cell death. Cells were transduced with Ad-PrxⅢ or Ad-Stuffer for 24 h and then exposed to 1 μM DOX for 18 h. (A) Cell lysates were subjected to immunoblot analyses for cleaved Caspase-3 and cleaved PARP-1 with normalization to β-actin. Densitometric quantification of protein expression was performed from three independent biological replicates. (B) Apoptotic cell death was assessed by Annexin V-FITC and 7-AAD double staining followed by flow cytometric analysis. Quantification of Annexin V and/or 7-AAD-positive cells is shown. All data are presented as mean ± S.D. Statistical significance was determined using two-way ANOVA followed by Bonferroni's post hoc test. ∗p < 0.05, ∗∗p < 0.01 and ∗∗∗p < 0.001.

Article Snippet: Rat cardiomyocyte cell line, H9c2 cells (CRL-1446) were obtained from the American Type Culture Collection (Manassas, VA, USA) and cultured in DMEM high glucose (SH30022.01, Hyclone, Logan, UT, USA) supplemented with 10% FBS, 1% antibiotic-antimycotic, and 1 μg/ml puromycin.

Techniques: Transduction, Western Blot, Expressing, Double Staining

Journal: Regenerative Therapy

Article Title: Lung cancer cell-derived exosomal EHF drives M2 macrophage polarization via transcriptional activation of RNF41 to promote tumor progression

doi: 10.1016/j.reth.2026.101104

Figure Lengend Snippet: EHF promotes lung cancer cell malignancy and M2 macrophage polarization. A549 and H520 cells were transfected with control (Ctrl) or EHF-knockdown (KD-EHF) plasmids. (A-B) Western blot and quantification of EHF protein levels in A549 and H520 cells. (C-D) EdU staining and quantification of EdU + cells in A549 and H520 cells, assessing proliferation. (E-H) Glucose uptake and lactate production assays were conducted using commercial kits to evaluate glycolytic output. (I-J) Flow cytometric analysis of apoptosis. (K-L) Transwell assay was employed to examine cell migration. (M − P) THP-1 cells were differentiated into THP-1-M0 macrophages with 100 ng/mL PMA for 24 h. Culture supernatants from Ctrl- or KD-EHF–transfected A549 and H520 cells were collected and co-incubated with THP-1-M0 macrophages. (M − N) Flow cytometric analysis was performed to assess the proportion of CD206 + M2-polarized macrophages. (O–P) ELISA was conducted to quantify the secretion of M2-associated cytokines (TGF-β1, IL-10, and VEGFA) in the supernatants of THP-1-M0 macrophages. Data are presented as mean ± SD. P < 0.05 denotes statistical significance.

Article Snippet: Human monocyte cell line (THP-1) (Catalog #: YC-D011) were purchased from Guangzhou Ubigene Biosciences Co., Ltd. (Guangzhou, China) and cultured in RPMI-1640 (Gibco) containing 10% FBS (Gibco).

Techniques: Transfection, Control, Knockdown, Western Blot, Staining, Transwell Assay, Migration, Incubation, Enzyme-linked Immunosorbent Assay

Journal: Regenerative Therapy

Article Title: Lung cancer cell-derived exosomal EHF drives M2 macrophage polarization via transcriptional activation of RNF41 to promote tumor progression

doi: 10.1016/j.reth.2026.101104

Figure Lengend Snippet: Exosomes derived from lung cancer cells are internalized by THP-1-M0 macrophages and mediate EHF delivery. Exosomes were isolated from A549 and H520 cells. (A) TEM images showed the typical cup-shaped morphology of exosomes isolated from A549 and H520 cell supernatants (scale bar: 100 nm). (B) NTA of exosomes from A549 and H520 cells. (C) Western blot analysis of exosome markers (CD9 and TSG101) and the endoplasmic reticulum marker Calnexin in exosome lysates and parental A549/H520 cell lysates. (D-E) THP-1-M0 macrophages (differentiated from THP-1 cells with 100 ng/mL PMA for 24 h) were co-incubated with DiI-labeled exosomes isolated from A549 and H520 cells. (D) Fluorescence microscopy images of THP-1-M0 macrophages after co-incubation with PKH67-labeled exosomes (scale bar: 20 μm). (E-F) Western blot and quantification of EHF protein levels in THP-1-M0 macrophages after incubation with exosomes from A549 or H520 cells. Data are presented as mean ± SD. P < 0.05 denotes statistical significance.

Article Snippet: Human monocyte cell line (THP-1) (Catalog #: YC-D011) were purchased from Guangzhou Ubigene Biosciences Co., Ltd. (Guangzhou, China) and cultured in RPMI-1640 (Gibco) containing 10% FBS (Gibco).

Techniques: Derivative Assay, Isolation, Western Blot, Marker, Incubation, Labeling, Fluorescence, Microscopy

Journal: Regenerative Therapy

Article Title: Lung cancer cell-derived exosomal EHF drives M2 macrophage polarization via transcriptional activation of RNF41 to promote tumor progression

doi: 10.1016/j.reth.2026.101104

Figure Lengend Snippet: Exosomal EHF reprograms THP-1-M0 macrophages to promote lung cancer cell malignancy and angiogenesis via paracrine signaling. (A) Schematic of the paracrine co-culture system: Exosomes from control (Ctrl) or EHF-knockdown (KD-EHF) A549/H520 cells were co-incubated with THP-1-M0 macrophages. The medium from these macrophages was used to treat A549/H520 cells or HUVECs. (B-E) THP-1-M0 macrophages were treated with three conditions: blank control (no exosome), exosomes from control-transfected A549/H520 cells (Exo Ctrl ), or exosomes from EHF-knockdown A549/H520 cells (Exo KD−EHF ). (B–C) Flow cytometry analysis of CD206 + M2 macrophage proportions in THP-1-M0 cells. (D-E) ELISA measurement of M2-associated cytokines (TGF-β1, IL-10, and VEGFA). (F-M) The medium from exosome treated macrophages was then used to incubate A549/H520 cells or HUVECs. (F-G) EdU staining and quantification of EdU + proliferating A549 and H520 cells. (H–I) Flow cytometric analysis of apoptosis in A549 and H520 cells. (J-K) Transwell migration assays and quantification of migrated A549 and H520 cells. (L-M) Tube formation assays and quantification of tube numbers in HUVECs. Data are presented as mean ± SD. P < 0.05 denotes statistical significance.

Article Snippet: Human monocyte cell line (THP-1) (Catalog #: YC-D011) were purchased from Guangzhou Ubigene Biosciences Co., Ltd. (Guangzhou, China) and cultured in RPMI-1640 (Gibco) containing 10% FBS (Gibco).

Techniques: Co-Culture Assay, Control, Knockdown, Incubation, Transfection, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Staining, Migration

Journal: Regenerative Therapy

Article Title: Lung cancer cell-derived exosomal EHF drives M2 macrophage polarization via transcriptional activation of RNF41 to promote tumor progression

doi: 10.1016/j.reth.2026.101104

Figure Lengend Snippet: EHF transcriptionally activates RNF41 and mediates its intercellular transfer via exosomes. (A) JASPAR database prediction of the EHF-binding motif within the RNF41 promoter region, localized to positions −828/-821. (B) ChIP-qPCR analysis in THP-1-M0 macrophages. (C) Dual-luciferase reporter assays in 293T cells. (D) WB analysis of EHF protein levels in THP-1-M0 macrophages with EHF knockdown (KD-EHF) or control (Ctrl). (E-F) Western blot and qRT-PCR analysis of RNF41 protein and mRNA levels in THP-1-M0 macrophages with KD-EHF or Ctrl. (G-H) THP-1-M0 macrophages were treated with culture medium (Blank), control exosomes (Ctrl Exo, from A549/H520 cells), or EHF-knockdown exosomes (KD-EHF Exo, from A549/H520-KD-EHF cells). RNF41 protein levels were detected by Western blot. Data are presented as mean ± SD. P < 0.05 denotes statistical significance.

Article Snippet: Human monocyte cell line (THP-1) (Catalog #: YC-D011) were purchased from Guangzhou Ubigene Biosciences Co., Ltd. (Guangzhou, China) and cultured in RPMI-1640 (Gibco) containing 10% FBS (Gibco).

Techniques: Binding Assay, ChIP-qPCR, Luciferase, Knockdown, Control, Western Blot, Quantitative RT-PCR

Journal: Regenerative Therapy

Article Title: Lung cancer cell-derived exosomal EHF drives M2 macrophage polarization via transcriptional activation of RNF41 to promote tumor progression

doi: 10.1016/j.reth.2026.101104

Figure Lengend Snippet: RNF41 mediates exosomal EHF-driven pro-tumorigenic effects in lung cancer. (A) Schematic of the rescue co-culture system: THP-1-M0 macrophages (control or RNF41-overexpressing) were incubated with exosomes from control (Exo Ctrl ) or EHF-knockdown (Exo KD−EHF ) A549/H520 cells. The medium from these co-cultures was used to treat A549/H520 cells or HUVECs. (B–C) Flow cytometry was used to analyze the proportion of CD206 + M2-polarized macrophages in THP-1-M0 cells. (D-E) ELISA was employed to quantify the levels of M2-associated cytokines (TGF-β1, IL-10, and VEGFA) in the CM from macrophages. (F-G) EdU assays were carried out to measure the proliferation of A549 and H520 cells. (H–I) Annexin V/PI flow cytometry was used to assess the apoptosis of A549 and H520 cells treated with CM. (J-K) Transwell assays were performed to evaluate the migration of A549 and H520 cells. (L-M) Tube formation assays were conducted on HUVECs treated with CM. Data are presented as mean ± SD. P < 0.05 denotes statistical significance.

Article Snippet: Human monocyte cell line (THP-1) (Catalog #: YC-D011) were purchased from Guangzhou Ubigene Biosciences Co., Ltd. (Guangzhou, China) and cultured in RPMI-1640 (Gibco) containing 10% FBS (Gibco).

Techniques: Co-Culture Assay, Control, Incubation, Knockdown, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Migration

Journal: The Journal of Biological Chemistry

Article Title: Improvement in binding and function of a monoclonal antibody against Shigella flexneri 3a O-antigen via phage display and whole-cell in-solution panning

doi: 10.1016/j.jbc.2026.111405

Figure Lengend Snippet: Functional analysis of hFlex3a2_v2 variants against S. flexneri 3a. A , L-ABA assay comparing the ability of single hFlex3a2_v2 variants or ( B ) combinatorial hFlex3a2_v2 variants and a non-binding control to coordinate complement-mediated bacteriolysis of S. flexneri 3a. L-ABA is the average of three biological replicates each performed in duplicate. L-ABAs in panels ( A ) and ( B ) were performed in parallel but graphed separately for clarity. hFlex3a2_v2 and control mAb data is the same in these two panels. p values in were calculated on logIC 50 values by repeated measure Anova with Dunnett’s correction for multiple comparisons. C , OPA assay measures the percentage of THP-1 cells that have phagocytosed S. flexneri 3a that was opsonized with 30 nM of the indicated mAb. This experiment is the result of five biological replicates each performed in duplicate, and p values were calculated by repeated measure ANOVA with Dunnett’s correction for multiple comparisons. Error bars represent standard deviation. D , Invasion assay shows the percent invasion of S. flexneri 3a incubated with 1 μM of the indicated mAb normalized to the invasion rate of S. flexneri 3a in PBS. This experiment is the result of four biological replicates each performed in duplicate, and p -values were calculated by repeated measure ANOVA with Dunnett’s correction for multiple comparisons. Error bars represent standard deviation. For all data, p values can be found in and those < 0.5 are denoted by a ∗. ( E ) A cartoon depicting the top-ranked Alphafold model of hFlex3a2_v2 (pLDDT = 96. 2, pTM = 0.929, and ipTM = 0.909). The VL is shown in light gray, and the VH is shown in dark gray. CDRL loops are colored orange, and residues where variants were selectively enriched against S. flexneri 3a following panning against S. flexneri 3a and S. sonnei and were experimentally characterized are colored green , while H34 (not selectively enriched) is shown in pink .

Article Snippet: THP-1 cell lines were obtained from ATCC.

Techniques: Functional Assay, Binding Assay, Control, OPA Assay, Standard Deviation, Invasion Assay, Incubation

Journal: The Journal of Biological Chemistry

Article Title: Improvement in binding and function of a monoclonal antibody against Shigella flexneri 3a O-antigen via phage display and whole-cell in-solution panning

doi: 10.1016/j.jbc.2026.111405

Figure Lengend Snippet: Functional analysis of hFlex3a2_v2 variants against S. flexneri 3b. Binding curves from a titration ELISA against S. flexneri 3b OMVs (2.5 μg/ml) show relative binding potencies of hFlex3a2_v2 ( A ) single amino acid variants or ( B ) hFlex3a2_v2 combinatorial amino acid variants. ELISA binding curves are the average of three independent replicates each performed in duplicate. ELISAs in panels ( A ) and ( B ) were performed in parallel but graphed separately for clarity. hFlex3a2_v2 and control mAb data is the same in these two panels. ( C ) L-ABA assay comparing the ability of single hFlex3a2_v2 variants or ( D ) combinatorial hFlex3a2_v2 variants and a non-binding control to coordinate complement-mediated bacteriolysis of S. flexneri 3b. L-ABA is the average of three biological replicates each performed in duplicate. L-ABAs in panels ( C ) and ( D ) were performed in parallel but graphed separately for clarity. hFlex3a2_v2 and control mAb data is the same in these two panels. p -values were calculated on logIC 50 values by repeated measure Anova with Dunnett’s correction for multiple comparisons ( E ) OPA assay measures the percent of THP-1 cells that have phagocytosed S. flexneri 3b that was opsonized with 30 nM of the indicated mAb. This experiment is the result of five biological replicates each performed in duplicate, and p -values were calculated by repeated measure ANOVA with Dunnett’s correction for multiple comparisons. Error bars represent standard deviation. F , invasion assay shows the percent invasion of S. flexneri 3b incubated with 1 μM of the indicated mAb normalized to the invasion rate of S. flexneri 3b in PBS. This experiment is the result of four biological replicates each performed in duplicate. Error bars represent standard deviation. For all plots, p values can be found in and those < 0.5 are denoted by a ∗.

Article Snippet: THP-1 cell lines were obtained from ATCC.

Techniques: Functional Assay, Binding Assay, Titration, Enzyme-linked Immunosorbent Assay, Control, OPA Assay, Standard Deviation, Invasion Assay, Incubation

Journal: International Journal of Molecular Medicine

Article Title: ZEB1 maintains mitochondrial fission and macrophage efferocytosis by restraining MFN2, thereby limiting inflammation and improving tendon-bone healing

doi: 10.3892/ijmm.2026.5805

Figure Lengend Snippet: ZEB1 knockdown affects the cell efferocytosis efficiency of macrophages. (A) After 6 days of induction culture with M-CSF, the expression level of F4/80 (green) was detected by immunofluorescence staining (scale bar, 20 µ m). (B) Flow cytometry was performed to detect the proportion of F4/80-positive cells to verify the purity of macrophages. (C) The mRNA expression levels of ZEB1 in macrophages at different time points (0, 1, 3 and 6 h) of co-culture were determined by RT-qPCR. (D) After co-incubation of BMDMs with ACs for 0, 1, 3 and 6 h, the protein expression of ZEB1 was detected by western blotting and semi-quantitative analysis of ZEB1 expression was conducted. (E) The mRNA expressions of TNFα, iNOS, CD206 and Arg-1 were detected by RT-qPCR. (F) The expression of ZEB1 mRNA was detected by RT-qPCR to evaluate the knockout efficiency. (G) Detection of ZEB1 protein expression by WB and semi-quantitative analysis of ZEB1 expression. (H) BMDMs were labeled with Mito-Tracker (green label), and then co-cultured with PI-labeled apoptotic Jurkat cells at a ratio of 5:1 of apoptotic cells to macrophages for 6 h. After removing the non-phagocytic apoptotic cells, the proportion of PI-positive macrophages (indicated by arrows) among the total macrophages was determined by fluorescence microscopy (Scale bar, 20 µ m). (I) The apoptosis rate of cells induced by UV radiation was detected by flow cytometry (Annexin V-FITC/PI double staining) to prepare target cells for the efferocytosis assay. n=3. * P<0.05 vs. NC or 0 h groups. NC, negative control; sh, short hairpin; ZEB1, zinc finger E-box binding homeobox 1; BMDMs, bone marrow-derived macrophages; ACs, apoptotic cells; M-CSF, macrophage colony-stimulating factor; RT-qPCR, reverse transcription-quantitative PCR; iNOS, inducible nitric oxide synthase.

Article Snippet: The human lymphocyte line Jurkat (clone E6-1; American Type Culture Collection) was selected as the apoptotic cell model. Logarithmic growth phase Jurkat cells (1×10 6 cells/ml) were seeded in six-well plates and exposed to ultraviolet (UV) light under uncovered conditions.

Techniques: Knockdown, Expressing, Immunofluorescence, Staining, Flow Cytometry, Co-Culture Assay, Quantitative RT-PCR, Incubation, Western Blot, Knock-Out, Labeling, Cell Culture, Fluorescence, Microscopy, Double Staining, Negative Control, Binding Assay, Derivative Assay, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Cancer Diagnosis & Prognosis

Article Title: Elevated Protein Tyrosine Phosphatase Kappa Expression Is Associated With Disease Progression and Poor Prognosis of Pancreatic Cancer

doi: 10.21873/cdp.10560

Figure Lengend Snippet: Verification of protein tyrosine phosphatase kappa (PTPRK) knockdown in pancreatic cancer cell lines. (A) QPCR results show the PTPRK expression in control cell line PANC-1 pEF and PTPRK knockdown cell line PANC-1 PTPRK kd . (B) PTPRK expression in CFPAC-1 pEF and CFPAC-1 PTPRK kd cell lines. (C) Western blot results show the PTPRK protein expression in both PANC-1 and CFPAC-1 cell lines with PTPRK nockdown. * p <0.05, ** p <0.01, *** p <0.001.

Article Snippet: Human pancreatic cancer cell lines PANC-1 and CFPAC-1 were purchased from ATCC (American Type Culture Collection, Manassas, VA, USA).

Techniques: Knockdown, Expressing, Control, Western Blot

Journal: Cancer Diagnosis & Prognosis

Article Title: Elevated Protein Tyrosine Phosphatase Kappa Expression Is Associated With Disease Progression and Poor Prognosis of Pancreatic Cancer

doi: 10.21873/cdp.10560

Figure Lengend Snippet: Protein tyrosine phosphatase kappa (PTPRK) and cell proliferation. (A, B) A proliferation was performed to examine whether PTPRK is associated with pancreatic cell proliferation. (C, D) QPCR results show the expression of CDK6 and CCND1 in control cell lines PANC-1 pEF /CFPAC-1 pEF and PTPRK knockdown cell lines PANC-1 PTPRK kd /CFPAC-1 PTPRK kd . (E) TCGA dataset is used to draw a scatter plot showing the association between CDK6 and PTPRK at transcripts level. (F) In the TCGA dataset, the association between CCND1 and PTPRK transcript levels is shown. * p <0.05, ** p <0.01, *** p <0.001.

Article Snippet: Human pancreatic cancer cell lines PANC-1 and CFPAC-1 were purchased from ATCC (American Type Culture Collection, Manassas, VA, USA).

Techniques: Expressing, Control, Knockdown

Journal: Cancer Diagnosis & Prognosis

Article Title: Elevated Protein Tyrosine Phosphatase Kappa Expression Is Associated With Disease Progression and Poor Prognosis of Pancreatic Cancer

doi: 10.21873/cdp.10560

Figure Lengend Snippet: Response to cyclin-dependent kinase 6 (CDK6) inhibitors in the protein tyrosine phosphatase kappa (PTPRK) knockdown pancreatic cancer cell line models. Both CFPAC-1 and PANC-1 cell lines were treated with different concentration of the CDK6 inhibitor BSJ-03-123 (A and B), CDK4/6 inhibitor Palbociclib (C and D) and CDK4 inhibitor 3-ATA (E and F). Corresponding IC 50 test results are shown. Cell viability was determined following a 3-day treatment with the inhibitors. * p <0.05, ** p <0.01, *** p <0.001.

Article Snippet: Human pancreatic cancer cell lines PANC-1 and CFPAC-1 were purchased from ATCC (American Type Culture Collection, Manassas, VA, USA).

Techniques: Knockdown, Concentration Assay

Journal: Cancer Diagnosis & Prognosis

Article Title: Elevated Protein Tyrosine Phosphatase Kappa Expression Is Associated With Disease Progression and Poor Prognosis of Pancreatic Cancer

doi: 10.21873/cdp.10560

Figure Lengend Snippet: Protein tyrosine phosphatase kappa (PTPRK) and lymph node metastasis. (A) The scatter plot shows that the lymph angiogenesis marker VEGFC is inversely correlated with PTPRK in the TCGA cohort. QPCR shows the expression of VEGFC in pancreatic cancer cell lines PANC-1 (B) and CFPAC-1 (C) with PTPRK knockdown. * p <0.05, ** p <0.01, *** p <0.001.

Article Snippet: Human pancreatic cancer cell lines PANC-1 and CFPAC-1 were purchased from ATCC (American Type Culture Collection, Manassas, VA, USA).

Techniques: Marker, Expressing, Knockdown

Journal: Translational Oncology

Article Title: Unmasking FCGR2B as a high-grade serous ovarian cancer specific marker of immune suppression and tumor progression through multi-omics mining

doi: 10.1016/j.tranon.2026.102748

Figure Lengend Snippet: FCGR2B expression defines an immunosuppressive TME in HGSOC. (A) ssGSEA-based immune infiltration analysis across 28 immune cell types. Δ = High - Low. (B) CIBERSORT-based macrophage-focused immune deconvolution reveals distinct immune landscapes between FCGR2B -High and FCGR2B -Low tumors. (C) Fluorescence images (scale bar, 50 μm) of FCGR2B (red), CD14 (green), CD68 (yellow), and the corresponding merged image. (D-E) Quantitative analysis of FCGR2B co-localization with CD14 (D) and CD68 (E). Left: box plots comparing co-localization levels between FCGR2B-High and FCGR2B-Low tumors (defined by the median FCGR2B H-score; Wilcoxon rank-sum test). Right: scatter plots showing correlations between FCGR2B and marker H-scores across all HGSOC samples (Spearman’s rho). (F) FCGR2B protein expression in HGSOC cell lines (SKOV3 and OVCAR3) and the cell line THP-1, as detected by WB assays. (G) FCGR2B protein expression in THP-1 cells transfected with Si-FCGR2B or control siRNA, as detected by WB assays. (H) Effect of FCGR2B knockdown on macrophage M2 polarization (CD206⁺/CD68⁺) under SKOV3 (HGSOC) co-culture conditions, assessed by flow cytometry. (I) Effect of FCGR2B knockdown on macrophage phagocytosis (CD68⁺/CFSE⁺) of SKOV3 cells under co-culture conditions, analyzed by flow cytometry. Statistical significance was determined using a two-tailed unpaired Student’s t -test; exact P values are indicated within the figure ( P = 0.0001, t = 14.35 for H; P = 0.0013, t = 7.976 for I; n =3, df = 4).

Article Snippet: The HGSOC cell lines SK-OV-3 (ATCC® HTB-77TM) and OVCAR-3 (ATCC® HTB-161TM), as well as the human monocytic cell line THP-1 (ATCC® TIB-202TM), were obtained from the American Type Culture Collection and cultured under standard conditions at 37 °C with 5% CO2.

Techniques: Expressing, Fluorescence, Marker, Transfection, Control, Knockdown, Co-Culture Assay, Flow Cytometry, Two Tailed Test