Journal: bioRxiv

Article Title: Salmonella -vectored vaccine delivering three Clostridium perfringens antigens protects poultry against necrotic enteritis

doi: 10.1101/318469

Figure Lengend Snippet: Adherence of C. perfringens CP4 to Caco-2 cells in the presence or absence of purified rFba, rNetB or rPspA. Attachment results significantly different from the no protein controls are indicated.

Article Snippet: The human colon carcinoma cell line Caco-2 (ATCC®#HTB-37) were obtained from the American Type Culture Collection (Manassas, VA) and cultured in Dulbecco’s’ modified Eagle’s medium (DMEM) with 4.5 g/L glucose (Corning, Manassas, VA) containing 4mM L-glutamine, 1% sodium pyruvate, 1% non-essential amino acids (NEEA), 100 U/ml penicillin, 100 μg/ml streptomycin, 20% heat inactivated fetal calf serum.

Techniques: Purification

Journal:

Article Title: LPP, an Actin Cytoskeleton Protein Related to Zyxin, Harbors a Nuclear Export Signal and Transcriptional Activation Capacity

doi:

Figure Lengend Snippet: Production of LPP in cells. (A) Scheme of the subdomain structure of the human zyxin, LPP, and TRIP6 proteins. All three proteins are composed of an N-terminal proline-rich domain (white box) and a C-terminal cysteine-rich portion composed of three LIM domains (gray boxes). LPP and zyxin contain FPPPP motifs in their N-terminal proline-rich domain (small boxes). (B) Specificity of anti-LPP antibodies. Total HeLa cell extracts were analyzed by SDS-PAGE and Western blotting with MP2, a serum directed against a synthetic LPP-derived peptide not present in zyxin or TRIP6 (amino acids 318–332; lane 1), with LPP2, an affinity-purified polyclonal antibody directed against a recombinant fusion protein containing amino acids 3–414 of LPP (GST-LPP3–414; lane 2), or with a monoclonal anti-zyxin antibody (lane 3). Note that zyxin migrates slower than LPP, confirming that LPP-specific antibodies do not recognize zyxin. The positions of molecular mass markers (kilodaltons) are shown on the left. (C) Expression of LPP in different human cells. Cell extracts were prepared from the following human cells and cell lines: foreskin fibroblasts (HFF), cervix carcinoma (HeLa), keratinocytes (Kera.), platelets (plat.), lymphocytes (HL60), and colon carcinoma (CaCo2). Approximately 15 μg of protein from each extract was analyzed by SDS-PAGE and Western blotting with MP2. The positions of molecular mass markers (kilodaltons) are shown on the left. (D) Quantitation of LPP and zyxin proteins in extracts from HFF, CaCo2, and LLC-PK1 cell lines. Values were obtained by quantifying LPP- or zyxin-specific protein bands with the use of a Cy5-based detection method (see MATERIALS AND METHODS). aThese values have been published elsewhere (Fradelizi et al., 1999 ) but are shown here for comparison.

Article Snippet: Cell Lines and Cells The human cervix carcinoma HeLa cell line (American Type Culture Collection [ATCC; Rockville, MD] CCL-2), African green monkey kidney Vero cell line (ATCC CCL-81), porcine proximal kidney epithelial LLC-PK1 cell line (ATCC CL-101), human colon adenocarcinoma CaCo2 cell line (ATCC HTB-37), human penis carcinoma keratinocytes ( Rogel-Gaillard et al. , 1992 ; Robine et al. , 1993 ), and human foreskin fibroblasts (HFF) (Dr. A. Rochat, Ecole Normale Superieure, Paris, France) were grown in DMEM supplemented with 10% FCS (complete medium).

Techniques: SDS Page, Western Blot, Derivative Assay, Affinity Purification, Recombinant, Expressing, Quantitation Assay, Comparison

Journal:

Article Title: LPP, an Actin Cytoskeleton Protein Related to Zyxin, Harbors a Nuclear Export Signal and Transcriptional Activation Capacity

doi:

Figure Lengend Snippet: Intracellular distribution of LPP in cultured cells. Cells were fixed, detergent permeabilized, and doubly stained by indirect immunofluorescence. (A–D) HFF cells doubly stained with affinity-purified LPP2 antibody (A and C) and with either monoclonal anti-vinculin (B) or anti-zyxin (D) antibody. Views were obtained by epifluorescence microscopy. The insets show 2.2-fold magnified details of images C and D. Ventral faces of the cells are shown. (E and F) Keratinocytes doubly stained with affinity-purified LPP2 antibody (E) and with monoclonal anti-vinculin antibody (F). Views were obtained by confocal laser scanning microscopy. Three apical sections were compiled. LPP codistributes with vinculin, which was used as a marker for focal adhesions (B) and cell-to-cell contacts (F). Note that in contrast to LPP, zyxin staining is not restricted to focal adhesions but is also found along actin stress fibers (D). LPP2 staining is also visible in the nucleus. Bars, 20 μm.

Article Snippet: Cell Lines and Cells The human cervix carcinoma HeLa cell line (American Type Culture Collection [ATCC; Rockville, MD] CCL-2), African green monkey kidney Vero cell line (ATCC CCL-81), porcine proximal kidney epithelial LLC-PK1 cell line (ATCC CL-101), human colon adenocarcinoma CaCo2 cell line (ATCC HTB-37), human penis carcinoma keratinocytes ( Rogel-Gaillard et al. , 1992 ; Robine et al. , 1993 ), and human foreskin fibroblasts (HFF) (Dr. A. Rochat, Ecole Normale Superieure, Paris, France) were grown in DMEM supplemented with 10% FCS (complete medium).

Techniques: Cell Culture, Staining, Immunofluorescence, Affinity Purification, Epifluorescence Microscopy, Confocal Laser Scanning Microscopy, Marker

Journal: Frontiers in Veterinary Science

Article Title: Botanicals as a zinc oxide alternative to protect intestinal cells from an Escherichia coli F4 infection in vitro by modulation of enterocyte inflammatory response and bacterial virulence

doi: 10.3389/fvets.2023.1141561

Figure Lengend Snippet: TER of Caco-2 cells treated with essential oils or powder extracts and challenged with E. coli F4 at 2 h (A) and 4 h (B) after the beginning of the infection. Groups with bacterial infection are represented with a “+” in the name. Data in the graphs are represented as means ± SEM; percentage values are referred to negative control (CTR–) for both the investigated timepoints. One-Way ANOVA analysis is performed against positive control (CTR+), identified with a red arrow; asterisks “*” denote significant differences with p < 0.05, while tendencies are highlighted by their p -value.

Article Snippet: The human colon carcinoma cell line (Caco-2) was obtained from DSMZ (Braunschweig, Germany).

Techniques: Infection, Negative Control, Positive Control

Journal: Frontiers in Veterinary Science

Article Title: Botanicals as a zinc oxide alternative to protect intestinal cells from an Escherichia coli F4 infection in vitro by modulation of enterocyte inflammatory response and bacterial virulence

doi: 10.3389/fvets.2023.1141561

Figure Lengend Snippet: Bacterial translocation of E. coli F4 across Caco-2 cells treated with essential oils or powder extracts at 2 h (A) and 4 h (B) after the beginning of the bacterial infection. Groups with bacterial infection are represented with a “+” in the name. Data in the graphs are represented as means ± SEM; percentage values are referred to positive control (CTR+) for both the investigated timepoints. One-Way ANOVA analysis is performed against positive control (CTR+), identified with a red arrow; asterisks “*” denote significant differences with p < 0.05, while tendencies are highlighted by their p -value.

Article Snippet: The human colon carcinoma cell line (Caco-2) was obtained from DSMZ (Braunschweig, Germany).

Techniques: Translocation Assay, Infection, Positive Control

Journal: Frontiers in Veterinary Science

Article Title: Botanicals as a zinc oxide alternative to protect intestinal cells from an Escherichia coli F4 infection in vitro by modulation of enterocyte inflammatory response and bacterial virulence

doi: 10.3389/fvets.2023.1141561

Figure Lengend Snippet: Gene expression analysis of Caco-2 cells treated with essential oils or powder extracts and challenged with E. coli F4 at 4 h after the beginning of the infection. The analyzed markers of cellular monolayer integrity are ZO-1 (A) , ZO-2 (B) , and CLD-1 (C) . Groups with bacterial infection are represented with a “+” in the name. Data in the graphs are represented as means ± SEM. One-Way ANOVA analysis is performed against positive control (CTR+), identified with a red arrow; asterisks “*” denote significant differences with p < 0.05, while tendencies are highlighted by their p -value.

Article Snippet: The human colon carcinoma cell line (Caco-2) was obtained from DSMZ (Braunschweig, Germany).

Techniques: Gene Expression, Infection, Positive Control

Journal: Frontiers in Veterinary Science

Article Title: Botanicals as a zinc oxide alternative to protect intestinal cells from an Escherichia coli F4 infection in vitro by modulation of enterocyte inflammatory response and bacterial virulence

doi: 10.3389/fvets.2023.1141561

Figure Lengend Snippet: Gene expression analysis of Caco-2 cells treated with essential oils or powder extracts and challenged with E. coli F4 at 4 h after the beginning of the infection. The analyzed markers of innate immune response are TNFα (A) , IL-1β (B) , IL-8 (C) , and BD1 (D) . Groups with bacterial infection are represented with a “+” in the name. Data in the graphs are represented as means ± SEM. One-Way ANOVA analysis is performed against positive control (CTR+), identified with a red arrow; asterisks “*” denote significant differences with p < 0.05, while tendencies are highlighted by their p -value.

Article Snippet: The human colon carcinoma cell line (Caco-2) was obtained from DSMZ (Braunschweig, Germany).

Techniques: Gene Expression, Infection, Positive Control

Journal: Frontiers in Veterinary Science

Article Title: Botanicals as a zinc oxide alternative to protect intestinal cells from an Escherichia coli F4 infection in vitro by modulation of enterocyte inflammatory response and bacterial virulence

doi: 10.3389/fvets.2023.1141561

Figure Lengend Snippet: Escherichia coli F4 adhesion assay performed on Caco-2 cells treated with essential oils or powder extracts for 1 h. Groups with bacterial infection are represented with a “+” in the name. Data in the graphs are represented as means ± SEM. One-Way ANOVA analysis is performed against positive control (CTR+), identified with a red arrow; asterisks “*” denote significant differences with p < 0.05.

Article Snippet: The human colon carcinoma cell line (Caco-2) was obtained from DSMZ (Braunschweig, Germany).

Techniques: Cell Adhesion Assay, Infection, Positive Control

Journal: Frontiers in Veterinary Science

Article Title: Botanicals as a zinc oxide alternative to protect intestinal cells from an Escherichia coli F4 infection in vitro by modulation of enterocyte inflammatory response and bacterial virulence

doi: 10.3389/fvets.2023.1141561

Figure Lengend Snippet: Immunofluorescence staining of Caco-2 cells treated with essential oils or powder extracts and simultaneously challenged with E. coli F4 for 2 h. Groups with bacterial infection are represented with a “+” in the name. In the image, the left column represents DAPI staining, the central column ZO-1 staining with FITC, while right column depicts the merge of the first two images, where white arrows identify areas of tight-junction detachment, loss of cells, holes in the monolayer or anomalies in ZO-1 disposition. Each row displays a different treatment.

Article Snippet: The human colon carcinoma cell line (Caco-2) was obtained from DSMZ (Braunschweig, Germany).

Techniques: Immunofluorescence, Staining, Infection

Journal: Frontiers in Veterinary Science

Article Title: Botanicals as a zinc oxide alternative to protect intestinal cells from an Escherichia coli F4 infection in vitro by modulation of enterocyte inflammatory response and bacterial virulence

doi: 10.3389/fvets.2023.1141561

Figure Lengend Snippet: Immunofluorescence staining of Caco-2 cells treated with essential oils or powder extracts and simultaneously challenged with E. coli F4 for 2 h. Groups with bacterial infection are represented with a “+” in the name. In the image, the left column represents DAPI staining, the central column ZO-1 staining with FITC, while right column depicts the merge of the first two images, where white arrows identify areas of tight-junction detachment, loss of cells, holes in the monolayer or anomalies in ZO-1 disposition. Each row displays a different treatment.

Article Snippet: The human colon carcinoma cell line (Caco-2) was obtained from DSMZ (Braunschweig, Germany).

Techniques: Immunofluorescence, Staining, Infection

Journal: Microbiology Spectrum

Article Title: Inhibition of protein kinase D and its substrate phosphatidylinositol-4 kinase III beta blocks common human coronavirus replication

doi: 10.1128/spectrum.01501-24

Figure Lengend Snippet: Infection by common human coronaviruses leads to an increased expression of PKD in cultured cells. ( A ) RT-qPCR was utilized to measure HCoV-229E viral RNA levels in MRC-5 cells, with results normalized to 18S RNA at various infection stages. ( B ) MRC-5 cells were infected with HCoV-229E at an MOI of 10 for a day, followed by 7 days of incubation period. Cell extracts were taken daily for RT-qPCR analysis (normalized to 18S RNA) and Western blotting (normalized to tubulin). Results are expressed as means ± SEM. ( C, D ) In a similar setup, MRC-5 cells were infected with HCoV-OC43 at an MOI of 10. ( E, F ) Another experiment was conducted using Caco-2-derived C2BBe1 cells infected with HcoV-NL63 at an MOI of 10. The results from these experiments represent data from three independent trials.

Article Snippet: C2BBe1 (clone of Caco-2) (human large intestine epithelial) (catalog no. CRL-2102) cells were purchased from ATCC.

Techniques: Infection, Expressing, Cell Culture, Quantitative RT-PCR, Incubation, Western Blot, Derivative Assay

Journal: Microbiology Spectrum

Article Title: Inhibition of protein kinase D and its substrate phosphatidylinositol-4 kinase III beta blocks common human coronavirus replication

doi: 10.1128/spectrum.01501-24

Figure Lengend Snippet: The inhibition of PKD3 attenuates common HCoV replication in cultured cells. ( A, B ) MRC-5 cells were transfected with PKD3 or control siRNAs for 48 h, after which RNA was extracted, and PKD levels were measured by RT-qPCR (normalized to GAPDH). Western blotting with antibodies for PKD3 and tubulin was also performed. ( C, D ) After infecting MRC-5 cells with HCoV-229E for 1 h, siRNAs were transfected for 48 h. The virus in the supernatants was quantified using RT-qPCR and plaque assays. ( E, F ) MRC-5 cells infected with HCoV-OC43 were analyzed similarly via RT-qPCR and plaque assays. ( G, H ) A similar experiment was performed in a C2BBe1 clone of Caco-2 cells infected with HCoV-NL63 and quantified by RT-qPCR and plaque assays. ( I, J ) The cells were infected with PKD3 adenovirus or control adenovirus for 48 h, after which RNA was extracted for PKD quantification by RT-qPCR (normalized to Gapdh), and Western blotting was performed. ( K, L ) After 1 h of HCoV-229E infection, MRC-5 cells were infected with PKD3 adenovirus or control adenovirus for 48 h, followed by viral quantification using RT-qPCR and plaque assays. ( M, N ) A similar process was applied for MRC-5 cells infected with HCoV-OC43. ( O, P ) C2BBe1 cells infected with HCoV-NL63 underwent a similar experiment for viral quantification.

Article Snippet: C2BBe1 (clone of Caco-2) (human large intestine epithelial) (catalog no. CRL-2102) cells were purchased from ATCC.

Techniques: Inhibition, Cell Culture, Transfection, Control, Quantitative RT-PCR, Western Blot, Virus, Infection

Journal: Microbiology Spectrum

Article Title: Inhibition of protein kinase D and its substrate phosphatidylinositol-4 kinase III beta blocks common human coronavirus replication

doi: 10.1128/spectrum.01501-24

Figure Lengend Snippet: The PKD pharmacological inhibitor CRT0066101 reduces common HCoV replication. ( A, B ) MRC-5 and C2BBe1 cells were exposed to increasing CRT0066101 concentrations for 24 h, and cell viability was measured by CCK8 assays. ( C, D ) MRC-5 cells infected with HCoV-229E (MOI of 10) were treated with increasing CRT0066101 concentrations for 72 h. Viral replication was assessed via TCID50 endpoint titer, RT-qPCR (normalized to 18S RNA), and plaque assays. (E) MRC-5 cells infected with HCoV-229E were treated with CRT0066101, fixed, immunostained using anti-TGN46 and anti-229E-RBD antibodies, and analyzed via confocal microscopy (bar = 10 µm). ( F-H ) Cells infected with HCoV-OC43 or HCoV-NL63 (MOI of 10) were treated with CRT0066101, and viral replication was measured by TCID50, RT-qPCR, and plaque assays. Graphs represent the means (±SEM) from three independent experiments, with statistical significance evaluated using one-way ANOVA followed by Dunnett’s post hoc test. *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Article Snippet: C2BBe1 (clone of Caco-2) (human large intestine epithelial) (catalog no. CRL-2102) cells were purchased from ATCC.

Techniques: Infection, Quantitative RT-PCR, Confocal Microscopy

Journal: Microbiology Spectrum

Article Title: Inhibition of protein kinase D and its substrate phosphatidylinositol-4 kinase III beta blocks common human coronavirus replication

doi: 10.1128/spectrum.01501-24

Figure Lengend Snippet: The PKD pharmacological inhibitor CRT0066101 blocks Golgi fission in the HCoV-infected cells. ( A ) MRC-5 cells infected with HcoV-229E and treated with CRT0066101 were immunostained using anti-TGN46 and anti-P230 antibodies, followed by confocal microscopy (bar = 10 µm). ( B, C ) The same process was repeated in MRC-5 and C2Bbe1 cells infected with HcoV-OC43 or HcoV-NL63 (bar = 10 µm).

Article Snippet: C2BBe1 (clone of Caco-2) (human large intestine epithelial) (catalog no. CRL-2102) cells were purchased from ATCC.

Techniques: Infection, Confocal Microscopy

Journal: Microbiology Spectrum

Article Title: Inhibition of protein kinase D and its substrate phosphatidylinositol-4 kinase III beta blocks common human coronavirus replication

doi: 10.1128/spectrum.01501-24

Figure Lengend Snippet: The PI4KIIIβ inhibitor BQR695 attenuates common HCoV replication and Golgi fission in cultured cells. ( A ) MRC-5 cells were treated with varying concentrations of BQR695 for 24 h, with cell viability measured by CCK8 assays. ( B, C ) HCoV-infected MRC-5 cells were treated with BQR695, fixed, and immunostained with anti-TGN46 and anti-P230 antibodies for confocal microscopy analysis (bar = 10 µm). ( D ) C2BBe1 clone of Caco-2 cells infected with HCoV-NL63 and then treated with BQR695, fixed, immunostained, and analyzed by confocal microscopy (bar = 10 µm). ( E-H ) MRC-5 and C2BBe1 cells infected with HCoV-229E, HCoV-OC43, or HCoV-NL63 were treated with BQR695, and viral replication was assessed by TCID50 and RT-qPCR. ( I-J ) Caco-2 cells, specifically the C2BBe1 clone, were infected with HCoV-NL63 at a multiplicity of infection (MOI) of 10. After allowing replication to proceed for 1 h, the cells were exposed to increasing concentrations of CRT0066101 for 24 h. *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Article Snippet: C2BBe1 (clone of Caco-2) (human large intestine epithelial) (catalog no. CRL-2102) cells were purchased from ATCC.

Techniques: Cell Culture, Infection, Confocal Microscopy, Quantitative RT-PCR

Journal: Cell Communication and Signaling : CCS

Article Title: Mitochondrial DNA release via mPTP and BAX/BAK drives inflammatory injury in intestinal ischemia reperfusion

doi: 10.1186/s12964-025-02603-3

Figure Lengend Snippet: A Representative transmission electron microscopy (TEM) images of mouse intestinal tissues and quantitative analysis of Swollen Mitochondria Ratio (SMR, ratio of swollen mitochondria to total mitochondria), Cristae Score, and crista density, n = 6, scale bar = 5 μm/500 nm. B qPCR analysis of the expression levels of mtDNA fragments 16S rRNA, ND1, and COX2 in mouse plasma, n = 6. C Representative images of TFAM immunohistochemical staining in murine intestinal tissues, n = 6. D Western blotting analysis of TFAM protein expression levels in mouse intestinal tissues, n = 6. E Western blotting analysis of TFAM protein expression levels in Caco-2 cells transduced with Ad-NC or Ad-TFAM, n = 6. F qPCR analysis of cytoplasmic mtDNA fragment ND1, D-LOOP levels in Caco-2 cells, n = 6. G Mice were intraperitoneally (ip) injected with solutions containing nDNA (5 mg/kg), mtDNA (5 mg/kg), or normal saline (NS), and plasma samples were collected 2 h post-injection. Levels of secreted IL-1β and IL-6 proteins in mouse plasma were analyzed by ELISA, n = 6. H qPCR analysis of the expression levels of mtDNA fragments ND1, D-LOOP, ATP6, and COX2 in Caco-2 cells after EtBr depletion, n = 6. I RT-qPCR analysis of IL-1β and IL-6 mRNA levels in Caco-2 cells, n = 6. Data are presented as mean ± SD. * P < 0.05

Article Snippet: The human colon carcinoma cell line Caco-2 was purchased from Wuhan Procell Life Technology Co., Ltd.

Techniques: Transmission Assay, Electron Microscopy, Expressing, Clinical Proteomics, Immunohistochemical staining, Staining, Western Blot, Transduction, Injection, Saline, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR

Journal: Cell Communication and Signaling : CCS

Article Title: Mitochondrial DNA release via mPTP and BAX/BAK drives inflammatory injury in intestinal ischemia reperfusion

doi: 10.1186/s12964-025-02603-3

Figure Lengend Snippet: Calcium overload leads to mPTP opening and subsequently causes mtDNA release following HR. A Representative fluorescent images of mPTP detection in Caco-2 cells via Calcein AM assay. Mito-Tracker Red CMXRos (red) is the mitochondrial internal reference, and green fluorescence reflects mPTP opening level, n = 6; bar = 50 μm. B Quantitative analysis of average mPTP fluorescence intensity (total regional fluorescence intensity/regional area) in each group via Image J, n = 6. C Image J was used for quantitative analysis of immunofluorescence colocalization. D qPCR analysis of the expression levels of cytosolic mtDNA fragments ND1, D-LOOP, ATP6, and COX2 in Caco-2 cells after treatment with CsA, n = 6. Data are presented as mean ± SD. * P < 0.05

Article Snippet: The human colon carcinoma cell line Caco-2 was purchased from Wuhan Procell Life Technology Co., Ltd.

Techniques: Calcein AM Assay, Fluorescence, Immunofluorescence, Expressing

Journal: Cell Communication and Signaling : CCS

Article Title: Mitochondrial DNA release via mPTP and BAX/BAK drives inflammatory injury in intestinal ischemia reperfusion

doi: 10.1186/s12964-025-02603-3

Figure Lengend Snippet: Mitochondrial calcium uniporter (MCU) mediated calcium overload regulates the opening of mPTP. A - C Western blotting ( A , B ) and RT-qPCR ( C ) analyses of MCU protein and mRNA expression levels in Caco-2 cells (si-MCU vs. si-NC), n = 6. D Representative immunofluorescence images of Rhod-2AM (calcium probe) and green mitochondrial fluorescent dye colocalization, and quantitative analysis using Image J, n = 6. E , H Representative immunofluorescence images of MCU (red), red mitochondrial fluorescent dye (green), and DAPI (nucleus, blue) ( E ), and quantitative analysis of MCU mitochondrial colocalization ( H ) in Caco-2 cells, n = 6, bar = 10 μm. F , I Representative immunofluorescence images of Rhod-2AM (red), green mitochondrial fluorescent dye (green), and Hoechst 33342 (blue) ( F ), and quantitative analysis of Rhod-2AM mitochondrial colocalization ( I ) in Caco-2 cells, n = 6, bar = 1 μm. G Image J-based quantitative analysis of immunofluorescence colocalization between MCU (red) and red mitochondrial fluorescent dye (green) in Caco-2 cells, n = 6. J , K Representative fluorescent images of mPTP detection (using calcein-AM/Co²⁺ probe) ( J ) and quantitative analysis of average fluorescence intensity ( K ) in Caco-2 cells, n = 6, bar = 50 μm. L Image J-based quantitative analysis of immunofluorescence colocalization between mPTP (labeled by calcein-AM, green) and red mitochondrial fluorescent dye (red) in Caco-2 cells, n = 6. M RT-qPCR analysis of cytosolic mtDNA fragment (ND1, D-LOOP, ATP6, COX2) expression levels in Caco-2 cells, n = 6.. Data are presented as mean ± SD. *P < 0.05

Article Snippet: The human colon carcinoma cell line Caco-2 was purchased from Wuhan Procell Life Technology Co., Ltd.

Techniques: Western Blot, Quantitative RT-PCR, Expressing, Immunofluorescence, Fluorescence, Labeling

Journal: Cell Communication and Signaling : CCS

Article Title: Mitochondrial DNA release via mPTP and BAX/BAK drives inflammatory injury in intestinal ischemia reperfusion

doi: 10.1186/s12964-025-02603-3

Figure Lengend Snippet: Oligomerization of BAX/BAK triggers the Mitochondrial Outer Membrane Permeabilization (MOMP) after HR. A - D Mitochondrial fractions were extracted through subcellular organelle separation. Western blotting analysis of the expression levels of monomeric and oligomeric forms of BAX and BAK proteins in mitochondria after BMH cross-linking treatment, n = 6. E - G Western blotting and RT-qPCR analysis of the protein and mRNA expression levels of BAX and BAK in Caco-2 cells, n = 6. H - K Mitochondrial fractions were extracted by subcellular separation. Western blotting analysis of the expression levels of monomeric and polymeric forms of BAX and BAK proteins in mitochondria after BMH cross-linking treatment, n = 6. L qPCR analysis of the expression levels of cytosolic mtDNA fragments ND1, D-LOOP, ATP6, and COX2 in Caco-2 cells, n = 6. Data are presented as mean ± SD. *P < 0.05

Article Snippet: The human colon carcinoma cell line Caco-2 was purchased from Wuhan Procell Life Technology Co., Ltd.

Techniques: Membrane, Western Blot, Expressing, Quantitative RT-PCR

Journal: Cell Communication and Signaling : CCS

Article Title: Mitochondrial DNA release via mPTP and BAX/BAK drives inflammatory injury in intestinal ischemia reperfusion

doi: 10.1186/s12964-025-02603-3

Figure Lengend Snippet: Independent but Sequential Activation of mPTP and BAX/BAK Leads to Release of mtDNA following HR. A Representative confocal immunofluorescence images and quantitative analysis of DNA, TOM20, and DAPI in Caco-2 cells, the number of cell= 10; bar = 10 μm. B Co-IP was performed using anti-CypD or anti-ANT1 antibodies, and BAX and BAK proteins were detected by Western blotting, n = 6. C Schematic diagram of the detection of mtDNA fragments in the IMS. D qPCR analysis of the expression levels of mtDNA fragments ND1, D-LOOP, ATP6, and COX2 in the IMS of Caco-2 cells, n = 3. E RT-qPCR analysis of the mRNA levels of IL-1β and IL-6 in Caco-2 cells, n = 6. Data are presented as mean ± SD. *P < 0.05

Article Snippet: The human colon carcinoma cell line Caco-2 was purchased from Wuhan Procell Life Technology Co., Ltd.

Techniques: Activation Assay, Immunofluorescence, Co-Immunoprecipitation Assay, Western Blot, Expressing, Quantitative RT-PCR