tlr7 polyclonal antibody  (Bioss)

Journal: iScience

Article Title: Evidence against the role of toll-like receptors 7 and 8 in sex selection in mice, cattle, and humans

doi: 10.1016/j.isci.2025.113164

Figure Lengend Snippet: Effects of TLR7/8 ligand (R848) on murine sperm partitioning and In vitro fertilization (IVF) (A) The schematic diagram of R848 treatment and swim-up layer assignment. (B) The percentages of sperm in different layers after treatment with R848 at various concentrations, ns = not significant ( p > 0.05). (C) The changes in the percentages of sperm of the same individual mice before and after R848 treatment in each swim-up layer. Data in (B and C) were analyzed using one-way ANOVA. (D and E) Cleavage (D) and blastocyst (E) rates of embryos derived from R848-treated sperm of different layers. Data were analyzed with one-way ANOVA followed by Tukey’s post-hoc tests. ∗ p ≤ 0.05, ∗∗ p ≤ 0.01. (F) The percentages of male embryos derived from IVF using sperm treated with 0 or 0.03 μM R848. The expected male embryo ratio was 50% (red bar). All values are mean ± SD of at least three replicates. (G) Representative agarose gel images of embryo sex determination by PCR. The product sizes for SRY and XIST were 105 and 147 bp, respectively. The bands below the specific ones were likely primer-dimers. Each lane represented an individual blastocyst. Lane marked with “F” and “M” were deemed to be female and male embryos, respectively.

Article Snippet: TLR7 polyclonal antibody , Bioss , Cat# bs-6601R; RRID:AB_11090775.

Techniques: In Vitro, Derivative Assay, Agarose Gel Electrophoresis

Journal: iScience

Article Title: Evidence against the role of toll-like receptors 7 and 8 in sex selection in mice, cattle, and humans

doi: 10.1016/j.isci.2025.113164

Figure Lengend Snippet: TLR7/8 ligand (R848) treatment did not separate X- and Y- sperm (A) Validation of TaqMan real-time PCR for sex chromosome ratio determination by using different ratios of male vs. female mouse genomic DNA. The expected ratios are marked with red bars. Data were analyzed with one sample t test. (B) TaqMan real-time PCR determination of the X-sperm ratios in different layers of sperm treated with R848. Data were analyzed with one-way ANOVA, ns = not significant ( p > 0.05). (C) Schematic diagram of centrifugation of sperm before R848 treatment and swim-up. (D) TaqMan real-time PCR determination of the X-sperm ratios in different layers of sperm from (C). Data were analyzed with one-way ANOVA. All values are mean ± SD from at least three replicates.

Article Snippet: TLR7 polyclonal antibody , Bioss , Cat# bs-6601R; RRID:AB_11090775.

Techniques: Biomarker Discovery, Real-time Polymerase Chain Reaction, Centrifugation

Journal: iScience

Article Title: Evidence against the role of toll-like receptors 7 and 8 in sex selection in mice, cattle, and humans

doi: 10.1016/j.isci.2025.113164

Figure Lengend Snippet: Localization of TLR7/8 in murine sperm (A) Immunofluorescence staining of TLR7/8 (green), acetyl α-tubulin (green), and DNA (blue) in murine caudal epididymal sperm. (i) The Bioss antibody against TLR7 stained the whole sperm tails; (ii) The Abcam antibody against TLR7 stained the lower half of the sperm tail as well as the acrosome of the sperm (red arrow); (iii) TLR8 antibody stained either the lower half (yellow arrow) or the entire sperm tail (blue arrow), as well as the acrosome (red arrow). Scale bar, 25 μm. (B) The percentages of murine sperm stained positively for TLR7/8 by immunofluorescence. (C) Flow cytometry analysis of mouse sperm stained with TLR7/8 and DAPI. In controls (left panels) the sperm were probed with the secondary antibody only and provided the cutoff (vertical lines in the upper panels) for stained and unstained sperm. The cells were detected by the FITC-A ( x axis) and DAPI ( y axis) channels. Each dot represents one sperm. The upper panel showed the distribution of the positively and negatively stained sperm, the lower panel displayed the intensity of fluorescence of the sperm ( x axis) and sperm counts ( y axis). The middle and right panels represent sperm stained for TLR7 and TLR8, respectively. Nearly all sperm were stained by both antibodies. (D) The percentages of mouse sperm positively stained for TLR7 or 8 as detected by flow cytometry. All values are mean ± SD of at least three replicates. Data were analyzed using t tests. ns = not significant ( p > 0.05).

Article Snippet: TLR7 polyclonal antibody , Bioss , Cat# bs-6601R; RRID:AB_11090775.

Techniques: Immunofluorescence, Staining, Flow Cytometry, Fluorescence

Journal: iScience

Article Title: Evidence against the role of toll-like receptors 7 and 8 in sex selection in mice, cattle, and humans

doi: 10.1016/j.isci.2025.113164

Figure Lengend Snippet: Detection of TLR7/8 on bovine X- and Y-sorted sperm (A) Immunofluorescence of TLR7 (green), TLR8 (green), acetyl α-tubulin (green) and DAPI (blue) on frozen-thawed un-sorted, X- and Y-sorted bovine sperm. (i–ii) Both TLR7 and TLR8 were stained at the acrosomal regions of the bovine sperm heads (red arrows), and tails (yellow arrows). Scale bar, 21 μm. (B, C, and E) (B) The percentages of positively stained bovine sperm for TLR7 and TLR8. Western blot analyses of TLR7 (C) and TLR8 (E) on bovine un-sorted (U), X-sorted (X) and Y-sorted (Y) sperm. TLR7 and TLR8 were resolved and blotted using non-reduced and reduced samples, respectively. (D and F) Quantification of the TLR7 (D) and TLR8 (F) bands from western blots. Data were analyzed using one-way ANOVA, ns = not significant ( p > 0.05). (G) Flow cytometry analysis of bovine sperm stained for TLR7/8 and DAPI. In controls (left panels) the sperm were probed with the secondary antibody only, which provided the cutoff (vertical line) for stained and unstained sperm. The cells were detected by the FITC-A ( x axis) and DAPI ( y axis) channels. Each dot represents one sperm. The middle and right panels represent sperm stained for TLR7 and TLR8, respectively. Nearly all sperm were stained by both antibodies. (H) The percentages of positively stained bovine sperm as detected by flow cytometry. Data were analyzed using one-way ANOVA. All values are mean ± SD of at least three replicates, ns = not significant ( p > 0.05).

Article Snippet: TLR7 polyclonal antibody , Bioss , Cat# bs-6601R; RRID:AB_11090775.

Techniques: Immunofluorescence, Staining, Western Blot, Flow Cytometry

Journal: iScience

Article Title: Evidence against the role of toll-like receptors 7 and 8 in sex selection in mice, cattle, and humans

doi: 10.1016/j.isci.2025.113164

Figure Lengend Snippet: Localization of TLR7 and TLR8 on human sperm (A) Representative images of immunofluorescence of TLR7/8 (green) and DAPI (blue) on human sperm. (i–ii) Both TLR7 and TLR8 stained human sperm tails as well as the equator regions of the heads (acrosomes, red arrows). TLR7 was also seen in the head-tail connecting apparatus (Yellow arrow). Scale bar, 22 μm. (B) The percentages of positively stained human sperm by TLR7/8 immunofluorescence. All values are mean ± SD of at least three replicates. Data were analyzed using t tests, ns = not significant ( p > 0.05). (C) Y chromosome fluorescence in situ hybridization (FISH, orange) and immunofluorescence for TLR7 and TLR8 (green) in a human sperm donor because his staining patterns and percentages were different from other donors. The inset (i) shows the details of TLR7 tail stain and the Y chromosome signal. TLR8 stained few sperm and the stain was seen on the entire sperm (ii). Immunofluorescence staining preceded FISH. Scale bar, 25 μm.

Article Snippet: TLR7 polyclonal antibody , Bioss , Cat# bs-6601R; RRID:AB_11090775.

Techniques: Immunofluorescence, Staining, Fluorescence, In Situ Hybridization

Journal: Biology

Article Title: Imiquimod (R837), a TLR7-Specific Agonist, Regulates Boar Sperm Motility via PI3K/GSK3α/β/Hexokinase Pathway

doi: 10.3390/biology14091182

Figure Lengend Snippet: The expression of TLR7 in boar sperm. ( A ) Localization and quantification of TLR7 (red) and DAPI (blue) in boar sperm. Sperm fluorescence scale bar = 50 μm. ( B ) Detection of TLR7 proteins in boar sperm by Western blotting.

Article Snippet: Sperm samples were placed on special glass slides and washed with PBS, air-dried, and permeabilized using TritonX-100/PBS (0.1% v / v ) for 1 h. Subsequently, the sperm sample was blocked with 10% goat serum ( v / v ) at room temperature for 30 min, followed by incubation with TLR7 primary antibody monoclonal (#bs 6601R, Bioss, 1:1000, Beijing, China) at 4 °C overnight.

Techniques: Expressing, Fluorescence, Western Blot

Journal: Biology

Article Title: Imiquimod (R837), a TLR7-Specific Agonist, Regulates Boar Sperm Motility via PI3K/GSK3α/β/Hexokinase Pathway

doi: 10.3390/biology14091182

Figure Lengend Snippet: Detection of TLR7 content in the upper layer and lower layer of boar sperm by FCM and WB. ( A , B ) The histogram of TLR7 expression was drawn based on flow cytometer detection. ( C ) The quantitative expression of TLR7 proteins divided by α-tubulin (internal control). ( D ) Gray analysis. Values are specified as mean ± standard error of the mean (SEM) of 4 replicates. Different lowercase letters indicate significant differences ( p < 0.05).

Article Snippet: Sperm samples were placed on special glass slides and washed with PBS, air-dried, and permeabilized using TritonX-100/PBS (0.1% v / v ) for 1 h. Subsequently, the sperm sample was blocked with 10% goat serum ( v / v ) at room temperature for 30 min, followed by incubation with TLR7 primary antibody monoclonal (#bs 6601R, Bioss, 1:1000, Beijing, China) at 4 °C overnight.

Techniques: Expressing, Flow Cytometry, Control

Journal: Biology

Article Title: Imiquimod (R837), a TLR7-Specific Agonist, Regulates Boar Sperm Motility via PI3K/GSK3α/β/Hexokinase Pathway

doi: 10.3390/biology14091182

Figure Lengend Snippet: The TLR7 signal transduction mechanism affects ATP production in boar sperm. In the lower-layer sperm, the TLR7 agonist R837 inhibits hexokinase activity and reduces ATP content by phosphorylation of PI3K and GSK3 proteins, thus leading to lower-layer sperm exhibiting low motility.

Article Snippet: Sperm samples were placed on special glass slides and washed with PBS, air-dried, and permeabilized using TritonX-100/PBS (0.1% v / v ) for 1 h. Subsequently, the sperm sample was blocked with 10% goat serum ( v / v ) at room temperature for 30 min, followed by incubation with TLR7 primary antibody monoclonal (#bs 6601R, Bioss, 1:1000, Beijing, China) at 4 °C overnight.

Techniques: Transduction, Activity Assay, Phospho-proteomics

Journal: International immunopharmacology

Article Title: Apigenin attenuated sepsis induced acute lung injury via polarizing macrophage towards M2 by blocking miR-146a →TLR7 interaction.

doi: 10.1016/j.intimp.2025.114446

Figure Lengend Snippet: Fig. 2. TLR7 signaling was enhanced in the lung and macrophage of septic mice. C57BL/6 J mice were intraperitoneally injected with LPS (10 mg/kg). Six h later, (A) TLR7 protein expression of mouse lungs treated with LPS (n = 3, Replication = 3) and (B) its quantization levels. (C) The phosphorylation level of p-ERK, p-p38 and p-JNK in the lungs of LPS-treated mice (n = 2, Replication = 3) and (D) their quantitative level. (E) TLR7 protein expression of mouse bone marrow-derived macrophages (BMDMs) treated with LPS (n = 3, Replication = 3) and (F) its quantization levels. (G) The phosphorylation level of p-ERK, p-p38 and p-JNK in the BMDMs of LPS-treated mice and (H) their quantitative level (n = 2, Replication = 3). (I) The phosphorylation level of p-ERK, p-p38 and p-JNK in Raw264.7 macrophage after 24 h of LPS (50 ng/ml) stimulation (n = 2, Replication = 3) and (J) their quantitative level. Data are presented as the means ± SEM and analyzed with unpaired t-test and two-way ANOVA. *p < 0.05, **p < 0.01, ***p < 0.001, ns, no significant.

Article Snippet: TLR7 polyclonal antibody (cat.17232–1-AP) was from Proteintech.

Techniques: Injection, Expressing, Phospho-proteomics, Derivative Assay

Journal: International immunopharmacology

Article Title: Apigenin attenuated sepsis induced acute lung injury via polarizing macrophage towards M2 by blocking miR-146a →TLR7 interaction.

doi: 10.1016/j.intimp.2025.114446

Figure Lengend Snippet: Fig. 3. TLR7 deficiency improves LPS induced pulmonary inflammation. (A) The mouse gene identification results after crossing C57BL/6 J mice with TLR7−/− mice showed that mice expressing the band at 305 bp but not at 404 bp are homozygous. WT mice (n = 4) and TLR7−/−mice (n = 5) were injected with LPS (10 mg/ kg), six h later, the blood and lungs were collected. (B) H&E staining of mouse lung pathology sections showing mice lung lesions. Scale bar, 50 μm. (C) The quantification of lung injuries. (D) Expression of mouse serum albumin at ELISA level and (E) expression of mouse serum IL-6. (F) mRNA expressions of IL-6 and TNF- α in mouse lungs. Data are presented as the means ± SEM and analyzed with two-way ANOVA. **p < 0.01, ***p < 0.001, ns, no significant.

Article Snippet: TLR7 polyclonal antibody (cat.17232–1-AP) was from Proteintech.

Techniques: Expressing, Injection, Staining, Enzyme-linked Immunosorbent Assay

Journal: International immunopharmacology

Article Title: Apigenin attenuated sepsis induced acute lung injury via polarizing macrophage towards M2 by blocking miR-146a →TLR7 interaction.

doi: 10.1016/j.intimp.2025.114446

Figure Lengend Snippet: Fig. 4. TLR7 deficiency increased macrophage alternative activation. WT mice (n = 3) and TLR7−/−(n = 6) mice were subjected to CLP. 24 h later, (A) the expression of macrophages in mouse BAL were detected by flow cytometry. (B) The total number of CD206 positive macrophage and its ratio in CD45 + cells were quantified. (C) Representative photomicrographs of pathological sections of the lungs labeled with CD206 and F4/80. From left to right, there are F4/80-specific labeling (green), CD206-specific labeling (pink), DAPI-stained cell nuclei (blue), and Merge plots of specific labeling and DAPI-stained cell nuclei. Scale bar, 20 μm. (D) Histograms showing changes in the phenotype of lentivirus-transfected Raw264.7 macrophages detected by flow cytometry. Data are presented as the means ± SEM and analyzed with unpaired t-test. *p < 0.05. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: TLR7 polyclonal antibody (cat.17232–1-AP) was from Proteintech.

Techniques: Activation Assay, Expressing, Flow Cytometry, Labeling, Staining, Transfection

Journal: International immunopharmacology

Article Title: Apigenin attenuated sepsis induced acute lung injury via polarizing macrophage towards M2 by blocking miR-146a →TLR7 interaction.

doi: 10.1016/j.intimp.2025.114446

Figure Lengend Snippet: Fig. 6. Api attenuated ALI and inflammation in a TLR7-dependent manner. WT mice (n = 6) and TLR7−/−mice (n = 6) were injected intraperitoneally with Api (30 mg/kg) for three consecutive days and then given a single dose (10 mg/kg) of LPS. Mice serum and lungs were collected after 6 h, (A) the phosphorylation level of p- ERK, p-p38 and p-JNK in the lungs of LPS-treated WT mice. (B) Expression of serum albumin and (C) IL-6 at ELISA level. (D) mRNA expressions of CCL2, IL-1β, IL-6 and TNF-α in mice lungs. (E) The phosphorylation level of p-ERK, p-p38 and p-JNK in the BMDMs of LPS-treated TLR7−/−mice. Data are presented as the means ± SEM and analyzed with two-way ANOVA. * p < 0.05, *** p < 0.001 and ns, no significant.

Article Snippet: TLR7 polyclonal antibody (cat.17232–1-AP) was from Proteintech.

Techniques: Injection, Phospho-proteomics, Expressing, Enzyme-linked Immunosorbent Assay

Journal: International immunopharmacology

Article Title: Apigenin attenuated sepsis induced acute lung injury via polarizing macrophage towards M2 by blocking miR-146a →TLR7 interaction.

doi: 10.1016/j.intimp.2025.114446

Figure Lengend Snippet: Fig. 7. Api improved sepsis induced ALI via inhibiting miR146a-TLR7 interaction in the macrophages. WT (n = 6) and TLR7−/− mice (n = 6) were given CLP surgery, injected with Api (30 mg/kg) one hour after surgery, and the mice were executed 24 h later. (A) mRNA expressions of miR-122, miR-146a, miR-let7 and miR-29a in mice lungs. (B, C) The miR-146a-5p mimic and inhibitor were introduced into Raw264.7 cells by liposome transfection. 24 h after transfection, LPS (50 ng/ml) and Api (10 μM) were added respectively, and the cells were collected 24 h later, and the expression of NOS2 was detected at the mRNA level. (D) LPS (50 ng/ ml) was added to lentivirus-transfected Raw264.7 cells, and the cells were collected after 24 h of stimulation to detect the mRNA expression level of miR-146a in macrophages. (E) RT-qPCR analysis of the efficiency of immunoprecipitation enrichment of RNA-binding protein-RNA complexes. Data are presented as the means ± SEM and analyzed with one-way ANOVA. ns, no significant, *p < 0.05, **p < 0.01 and ***p < 0.001.

Article Snippet: TLR7 polyclonal antibody (cat.17232–1-AP) was from Proteintech.

Techniques: Injection, Transfection, Expressing, Quantitative RT-PCR, Immunoprecipitation, RNA Binding Assay