anti romo 1  (Proteintech)

Journal: Respiratory Research

Article Title: ROMO1 is involved in airway mucus hypersecretion in COPD through the mitochondrial ROS-STAT6 pathway

doi: 10.1186/s12931-026-03578-6

Figure Lengend Snippet: Protein expression of ROMO1 and MUC5AC in the airway epithelium of control and COPD patients. Representative images of hematoxylin-eosin (HE) and immunohistochemistry (IHC) staining of lung tissues from the control and COPD patients ( A ). Quantitative analysis of the relative expression of ROMO1 ( B ) and MUC5AC ( C ) in the airway epithelium from the control and COPD patients. Data are presented as mean ± SD. * P < 0.05 compared with the control group

Article Snippet: The membranes were incubated with specific primary antibodies against ROMO1 (1:2000; OriGene Technology, USA), p-STAT6 (1:1000; Cell Signaling Technology, USA), STAT6 (1:1000; Santa Cruz Biotechnology, USA), and GAPDH (1:5000; Abcam, UK) at 4°C overnight, followed by incubation with the secondary antibodies for 1 h at room temperature.

Techniques: Expressing, Control, Immunohistochemistry

Journal: Respiratory Research

Article Title: ROMO1 is involved in airway mucus hypersecretion in COPD through the mitochondrial ROS-STAT6 pathway

doi: 10.1186/s12931-026-03578-6

Figure Lengend Snippet: ROMO1 and MUC5AC expression increased in a concentration-dependent manner in CSE-treated airway epithelial cells. The mRNA and protein expression of ROMO1 was increased when airway epithelial cells were treated with 2.5%-10% CSE for 24 h ( A - B ). Exposure to CSE (2.5%-10%) for 24 h increased MUC5AC mRNA expression in a concentration-dependent manner in the airway epithelial cells ( C ). Representative immunofluorescence images and their quantitative analysis demonstrated that MUC5AC expression was significantly elevated when airway epithelial cells were exposed to 2.5%-10% CSE for 24 h ( D - E ). The concentration of MUC5AC in the cell culture supernatant was significantly increased in a concentration-dependent manner under the same treatment condition ( F ). Experimental values are expressed as mean ± SD from three independent replicates. * P < 0.05 versus the control group

Article Snippet: The membranes were incubated with specific primary antibodies against ROMO1 (1:2000; OriGene Technology, USA), p-STAT6 (1:1000; Cell Signaling Technology, USA), STAT6 (1:1000; Santa Cruz Biotechnology, USA), and GAPDH (1:5000; Abcam, UK) at 4°C overnight, followed by incubation with the secondary antibodies for 1 h at room temperature.

Techniques: Expressing, Concentration Assay, Immunofluorescence, Cell Culture, Control

Journal: Respiratory Research

Article Title: ROMO1 is involved in airway mucus hypersecretion in COPD through the mitochondrial ROS-STAT6 pathway

doi: 10.1186/s12931-026-03578-6

Figure Lengend Snippet: CSE treatment induced a time-dependent increase in ROMO1 and MUC5AC expression in airway epithelial cells. The mRNA and protein expression of ROMO1 was significantly increased when airway epithelial cells were treated with 7.5% CSE for 8–32 h ( A - B ). A time-dependent increase in MUC5AC mRNA expression was observed in airway epithelial cells stimulated with 7.5% CSE for 8–32 h ( C ). Representative immunofluorescence images and quantitative analysis showed that treatment with 7.5% CSE induced a time-dependent up-regulation of MUC5AC protein expression in airway epithelial cells ( D - E ). The concentration of MUC5AC in cell culture supernatant was significantly increased following the treatment of airway epithelial cells with 7.5% CSE for 8–32 h ( F ). All statistical data are expressed as mean ± SD from three independent experiments. * P < 0.05 versus the 0 H group

Article Snippet: The membranes were incubated with specific primary antibodies against ROMO1 (1:2000; OriGene Technology, USA), p-STAT6 (1:1000; Cell Signaling Technology, USA), STAT6 (1:1000; Santa Cruz Biotechnology, USA), and GAPDH (1:5000; Abcam, UK) at 4°C overnight, followed by incubation with the secondary antibodies for 1 h at room temperature.

Techniques: Expressing, Immunofluorescence, Concentration Assay, Cell Culture

Journal: Respiratory Research

Article Title: ROMO1 is involved in airway mucus hypersecretion in COPD through the mitochondrial ROS-STAT6 pathway

doi: 10.1186/s12931-026-03578-6

Figure Lengend Snippet: ROMO1 silencing attenuated airway epithelial mitochondrial injury induced by CSE. Airway epithelial cells were transiently transfected with ROMO1 siRNA for 24 h, and the gene silencing effect was confirmed by western-blot ( A ). Real-time PCR ( B ) and western-blot ( C ) showed that ROMO1 knockdown significantly inhibited the increase of ROMO1 expression in airway epithelial cells treated with 7.5% CSE for 24 h. ROMO1 silencing significantly suppressed CSE-induced decline in cell viability ( D ). Transmission electron microscope examination revealed that pretreatment with ROMO1 siRNA attenuated airway epithelial mitochondrial structure damage induced by 7.5% CSE ( E ). Representative images of mitochondrial membrane potential (MMP) in airway epithelial cells are presented as determined by confocal microscope ( F ). Quantitative analysis showed that ROMO1 silencing significantly increased the levels of MMP ( G ) and intracellular ATP ( H ) in airway epithelial cells treated with 7.5% CSE for 24 h. Mitochondrial ROS content was determined by flow cytometry ( I ), and its level was significantly decreased in airway epithelial cells treated with ROMO1 siRNA and 7.5% CSE ( J ). All values are shown as mean ± SD from three independent experiments. * P < 0.05 versus the control group, and # P < 0.05 versus the 7.5% CSE group

Article Snippet: The membranes were incubated with specific primary antibodies against ROMO1 (1:2000; OriGene Technology, USA), p-STAT6 (1:1000; Cell Signaling Technology, USA), STAT6 (1:1000; Santa Cruz Biotechnology, USA), and GAPDH (1:5000; Abcam, UK) at 4°C overnight, followed by incubation with the secondary antibodies for 1 h at room temperature.

Techniques: Transfection, Western Blot, Real-time Polymerase Chain Reaction, Knockdown, Expressing, Transmission Assay, Microscopy, Membrane, Flow Cytometry, Control

Journal: Respiratory Research

Article Title: ROMO1 is involved in airway mucus hypersecretion in COPD through the mitochondrial ROS-STAT6 pathway

doi: 10.1186/s12931-026-03578-6

Figure Lengend Snippet: ROMO1 silencing decreased MUC5AC expression in airway epithelial cells treated with CSE. Real-time PCR ( A ) and fluorescence microscope ( B - C ) showed that ROMO1 knockdown significantly reduced MUC5AC expression in airway epithelial cells treated with 7.5% CSE for 24 h. The concentration of MUC5AC in the cell culture supernatant was significantly decreased following the treatment with ROMO1 siRNA and 7.5% CSE ( D ). Data are presented as mean ± SD from three independent experiments. * P < 0.05 versus the control group, and # P < 0.05 versus the 7.5% CSE group

Article Snippet: The membranes were incubated with specific primary antibodies against ROMO1 (1:2000; OriGene Technology, USA), p-STAT6 (1:1000; Cell Signaling Technology, USA), STAT6 (1:1000; Santa Cruz Biotechnology, USA), and GAPDH (1:5000; Abcam, UK) at 4°C overnight, followed by incubation with the secondary antibodies for 1 h at room temperature.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Fluorescence, Microscopy, Knockdown, Concentration Assay, Cell Culture, Control

Journal: Respiratory Research

Article Title: ROMO1 is involved in airway mucus hypersecretion in COPD through the mitochondrial ROS-STAT6 pathway

doi: 10.1186/s12931-026-03578-6

Figure Lengend Snippet: Mitochondrial ROS-STAT6 pathway mediated MUC5AC expression in airway epithelial cells exposed to CSE. Western-blot revealed that ROMO1 silencing significantly inhibited p-STAT6/STAT6 expression in airway epithelial cells treated with 7.5% CSE for 24 h ( A ). The protein expression of p-STAT6/STAT6 in 7.5% CSE-treated airway epithelial cells was significantly decreased when these cells were pretreated with MitoTEMPO or AS1517499 ( B ). Real-time PCR ( C ) and fluorescence microscopy ( D - E ) analysis demonstrated that pretreatment with either MitoTEMPO or AS1517499 significantly reduced MUC5AC expression in airway epithelial cells exposed to 7.5% CSE. The concentration of MUC5AC in the cell supernatant was significantly decreased under the same treatment conditions ( F ). Data are presented as mean ± SD from three independent experiments. * P < 0.05 versus the control group, and # P < 0.05 versus the 7.5% CSE group

Article Snippet: The membranes were incubated with specific primary antibodies against ROMO1 (1:2000; OriGene Technology, USA), p-STAT6 (1:1000; Cell Signaling Technology, USA), STAT6 (1:1000; Santa Cruz Biotechnology, USA), and GAPDH (1:5000; Abcam, UK) at 4°C overnight, followed by incubation with the secondary antibodies for 1 h at room temperature.

Techniques: Expressing, Western Blot, Real-time Polymerase Chain Reaction, Fluorescence, Microscopy, Concentration Assay, Control

Journal: Respiratory Research

Article Title: ROMO1 is involved in airway mucus hypersecretion in COPD through the mitochondrial ROS-STAT6 pathway

doi: 10.1186/s12931-026-03578-6

Figure Lengend Snippet: Schematic illustration of the mechanism of ROMO1 in airway mucus hypersecretion during COPD pathogenesis. ROMO1 up-regulation induces airway epithelial mitochondrial dysfunction, thereby contributing to airway mucus hypersecretion through the regulation of mitochondrial ROS-STAT6 signaling pathway in the pathogenesis of COPD

Article Snippet: The membranes were incubated with specific primary antibodies against ROMO1 (1:2000; OriGene Technology, USA), p-STAT6 (1:1000; Cell Signaling Technology, USA), STAT6 (1:1000; Santa Cruz Biotechnology, USA), and GAPDH (1:5000; Abcam, UK) at 4°C overnight, followed by incubation with the secondary antibodies for 1 h at room temperature.

Techniques:

Journal: JCI insight

Article Title: C3aR-initiated signaling is a critical mechanism of podocyte injury in membranous nephropathy.

doi: 10.1172/jci.insight.172976

Figure Lengend Snippet: Figure 3. Effect of anti-PLA2R serum on podocyte phenotype. (A and B) C3aR (A) and PLA2R1 (B) RNA expression in podocytes exposed to 0.5% healthy or MN serum for 72 hours. Red dotted line: hPOD in media with no serum, normalization group (n = 3/group). (C) Western Blotting bands for C3aR (54kDa) and PLA2R1 (150 kDa) and respective beta actin (42 kDa) exposed to healthy or anti-PLA2R serum (n = 3/group). (D) Western blotting analysis of C3aR (54kDa) in hPOD monolayers exposed to MN sera, confirming an increased expression after 72 hours. Bands were normalized against β-actin (42 kDa), showing a significantly increased expression. n = 3/group. (E) Western blotting analysis of PLA2R (150 kDa) in hPOD monolayers exposed to MN sera, showing a statis- tically significant increase in protein expression at 72 hours. Bands were normalized against β-actin (42 kDa), showing a significantly increased expression (n = 3/group). (F) Western Blotting bands for ROMO1 (10 kDa) and caspase-3 (35 kDa) and respective β-actin (42 kDa) exposed to healthy or anti-PLA2R serum (n = 3/group). (G) Western blotting analysis of oxidative stress, measured by expression of ROMO1 (10 kDa), in hPOD monolayers exposed to MN sera. Bands were normalized against β-actin (42 kDa), showing a significantly increased protein level (n = 3/group). (H) Western blotting analysis of apoptosis, measured by expression of caspase-3 (35 kDa), in hPOD monolayers exposed to MN sera. Bands were normalized against β-actin (42 kDa), showing an increased expression (n = 3/group). (I and J) hPOD monolayers exposed to healthy (I) or MN serum (J) confirmed an increase in ROMO1 (red). DAPI, blue. Scale bar: 25 µm. (K and L) Representative immunofluorescence staining of hPOD monolayers exposed to healthy (K) or MN serum (L) confirmed an increase in caspase-3 (red). DAPI, blue. Scale bar: 25 µm. All statistical values determined by 2-tailed Student’s t test. *P < 0.05, **P < 0.01.

Article Snippet: The following primary antibodies were used: PLA2R (LS-C108906-100, LSBio), C3AR (NBP2-15649, NovusBio), caspase-3 (9662S, Cell Signaling Technology), Synaptopodin (ab224491, Abcam), ROMO1 (NBP2-45607, NovusBio), C5b-9 (bs-2673R, Bioss), and THSD7A (HPA000923, Atlas Antibodies).

Techniques: RNA Expression, Western Blot, Expressing, Immunofluorescence, Staining

Journal: JCI insight

Article Title: C3aR-initiated signaling is a critical mechanism of podocyte injury in membranous nephropathy.

doi: 10.1172/jci.insight.172976

Figure Lengend Snippet: Figure 4. C3aR antagonism effectively prevents changes in podocyte phenotype by MN serum. (A) Western blotting analysis of C3aR in hPOD monolayers exposed to MN sera or MN sera + C3aRA. The antagonist efficient- ly prevented C3aR (54 kDa) increase. Bands were normalized against β-actin (42 KDa), showing a significantly decreased protein level. n = 3/group. (B) Western blotting analysis of PLA2R (150kDa) in hPOD monolayers exposed to MN sera or MN sera + C3aRA. The antagonist efficiently prevented PLA2R increase. Bands were normalized against β-actin (42 kDa), showing a significantly decreased protein level. n = 3/group. (C) Western blotting analysis of apoptosis, measured by expression of caspase-3 (35 kDa), in hPOD monolayers exposed to MN sera or MN sera + C3aRA. The antagonist significantly decreased apoptosis in hPOD. Bands were normalized against β-actin (42 kDa), showing a significantly decreased protein level. n = 3/group. (D) Western Blotting bands for Synaptopodin (100kDa) and ROMO1 (10 kDa) and respective β-actin (42 kDa) after exposure to healthy serum, anti-PLA2R serum, or anti-PLA2R serum + C3aRA. (n = 3/group). (E) Western blotting analysis of synaptopodin (100 kDa) in hPOD monolayers exposed to MN sera or MN sera+ C3aRA. The antagonist efficiently prevented synaptopodin loss. Bands were normalized against β-actin (42 kDa), showing a significantly increased protein level. Bands are shown in D. n of replicates/group: 3. (F) Western blotting analysis of oxidative stress, measured by expression of ROMO1 (10 kDa), in hPOD monolayers exposed to MN sera or MN sera+ C3aRA. The antagonist significantly decreased ROMO1 expression in hPOD. Bands were normalized against β-actin (42 kDa), showing a significantly decreased protein level. n = 3/group. Bands are shown in D. All statistical values determined by 1-way ANOVA. *P < 0.05; **P < 0.01, ***P < 0.001.

Article Snippet: The following primary antibodies were used: PLA2R (LS-C108906-100, LSBio), C3AR (NBP2-15649, NovusBio), caspase-3 (9662S, Cell Signaling Technology), Synaptopodin (ab224491, Abcam), ROMO1 (NBP2-45607, NovusBio), C5b-9 (bs-2673R, Bioss), and THSD7A (HPA000923, Atlas Antibodies).

Techniques: Western Blot, Expressing