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Santa Cruz Biotechnology sirna oligonucleotides
Sirna Oligonucleotides, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/il+10+sirna/IL-10+siRNA/pmc11865947-81-2-9
Average 93 stars, based on 14 article reviews

sirna oligonucleotides - by Bioz Stars, 2026-10

93/100 stars

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Knockdown:

Article Title: Role of lncRNA MAGI2-AS3 in lipopolysaccharide-induced nucleus pulposus cells injury by regulating miR-374b-5p/interleukin-10 axis.
Article Snippet: .. For IL‐10 knockdown, the control‐siRNA (sc‐36869) and IL‐10‐siRNA (sc‐39634) were purchased from Santa Cruz Biotechnology. .. NP cells were transfected with control‐plasmid, MAGI2‐AS3‐plasmid, control‐siRNA, IL‐10‐siRNA, mimic control (Shanghai GenePharma), miR‐374‐5p mimic (Shanghai GenePharma), inhibitor 2 of 13 | YU and LI 20504527, 2023, 4, D ow nloaded from https://onlinelibrary.w iley.com /doi/10.1002/iid3.772 by C ochrane Israel, W iley O nline L ibrary on [01/08/2023].

Article Title: Prostaglandin I2 (PGI 2 ) inhibits Brucella abortus internalization in macrophages via PGI 2 receptor signaling, and its analogue affects immune response and disease outcome in mice.
Article Snippet: To date, the implications of prostaglandin I2 (PGI2), a prominent lipid mediator for modulation of immune responses, has not been clearly understood in Brucella infection.. In this study, we found that cyclooxygenase-2 (COX-2) was significantly expressed in both infected bone marrow-derived macrophages (BMMs) and RAW 264.7 cells.. Prostaglandin I2 synthase (PTGIS) expression was not significantly changed, and PGI2 receptor (PTGIR) expression was downregulated in BMMs but upregulated in RAW 264.7 macrophages at late infection.

Control:

Article Title: Macrophage differentiation and polarization via phosphatidylinositol 3-kinase/Akt-ERK signaling pathway conferred by serum amyloid P component.
Article Snippet: .. RAW264.7 cells were transiently transfected with 100 nM control small interfering RNA (siRNA) or IL-10 siRNA (Santa Cruz Biotechnology, Santa Cruz, CA) using a Lipofectamine 2000 transfection reagent (Invitrogen Life Technologies) at 2:1 (lipid/siRNA) ratio following the manufacturer’s instructions for siRNA transfection. ..

Article Title: IL-6-mediated induction of MMP-9 is modulated by JAK-dependent IL-10 expression in macrophages
Article Snippet: Membranes were washed 2X in TBST and incubated 1 h in 3% blocking buffer containing goat anti-rabbit IgG conjugated to HRP (1:3000; BioRad Laboratories). .. IL-10 knockdowns RAW264.7 macrophages were transfected with control siRNA (i.e. nonspecific, NS) or IL-10 siRNA (Santa Cruz Biotechnology) utilizing HiPerFect transfection reagent according to the manufacturer's protocol (Qiagen). ..

Article Title: IL-6-mediated induction of MMP-9 is modulated by JAK-dependent IL-10 expression in macrophages
Article Snippet: Membranes were washed 2X in TBST and incubated 1 h in 3% blocking buffer containing goat anti-rabbit IgG conjugated to HRP (1:3000; BioRad Laboratories). .. RAW264.7 macrophages were transfected with control siRNA (i.e. nonspecific, NS) or IL-10 siRNA (Santa Cruz Biotechnology) utilizing HiPerFect transfection reagent according to the manufacturer's protocol (Qiagen). ..

Article Title: Chronic lymphocytic leukemia cells induce changes in gene expression of CD4 and CD8 T cells
Article Snippet: .. Relative IL-10 protein expression in CLL cells treated with nonspecific control siRNA or IL-10 siRNA was detected with anti–human IL-10 polyclonal antibody (Santa Cruz Biotechnology Inc.) and normalized to GAPDH protein level. ..

Concentration Assay:

Cell Culture:

Article Title: Carbon Monoxide Inhibits Tenascin-C Mediated Inflammation via IL-10 Expression in a Septic Mouse Model
Article Snippet: .. RAW 264.7 cells (5 × 10 5 /mL) were cultured in 6-well plates for 3 h and transfected with IL-10 siRNA (100 nM) or TN-C siRNA (100 nM) from Santa Cruz Biotechnology using Lipofectamine 2000 according to the manufacturer's instructions. ..

Transfection:

Article Title: IL-6-mediated induction of MMP-9 is modulated by JAK-dependent IL-10 expression in macrophages
Article Snippet: Membranes were washed 2X in TBST and incubated 1 h in 3% blocking buffer containing goat anti-rabbit IgG conjugated to HRP (1:3000; BioRad Laboratories). .. RAW264.7 macrophages were transfected with control siRNA (i.e. nonspecific, NS) or IL-10 siRNA (Santa Cruz Biotechnology) utilizing HiPerFect transfection reagent according to the manufacturer's protocol (Qiagen). ..

Small Interfering RNA:

Article Title: Carbon monoxide decreases interleukin-1β levels in the lung through the induction of pyrin
Article Snippet: Antibody against NLRP3 was purchased from IMGENEX (San Diego, CA, USA), and IL-10 was purchased from Novus Biologicals (Littleton, CO, USA). .. Scrambled small interfering RNA (siRNA), pyrin siRNA, and IL-10 siRNA were purchased from Santa Cruz Biotechnology. ..

Expressing:

Image Search Results

Mct4 expression is promoted by Mtb infection and exerts potent pro-Mtb effects among SLC family in macrophages (A) 5 × 10 5 of BMDMs in 12-well plates were extracted by Trizol method to conduct the subsequent experiment. Intracellular mRNA level was detected in BMDMs at 24 h of Mtb infection at MOI of 1 by RNA sequencing. (B) 5 × 10 5 of BMDMs in 12-well plates were extracted by Trizol method to conduct the subsequent experiment. Intracellular mRNA level was detected in BMDMs at 0, 6, 24, 48, and 72 h of Mtb infection at MOI of 1 by qRT-PCR analysis. β-Actin served as an internal control. (C) SLC gene-silenced BMDMs were detected for relative mRNA expression by quantitative reverse-transcription PCR (qRT-PCR) analysis. β-Actin served as an internal control. (D) The intracellular Mtb load was detected in SLC gene-silenced BMDMs at 0 and 48 h of Mtb infection at MOI of 5 by colony-forming unit (CFU) assay. (E) Intracellular mRNA level was detected in BMDMs at 0, 6, 24, 48, and 72 h of Mtb infection at MOI of 1 by qRT-PCR analysis. β-Actin served as an internal control. (F) Intracellular protein level was detected in BMDMs at 0, 6, 24, 48, and 72 h of Mtb infection at MOI of 1 by western blot analysis. The ratio of expression of Mct4 and β-actin is shown in graph. β-Actin served as an internal control. (G) Intracellular mRNA level was detected in Raw 264.7 at 0, 6, 24, 48, and 72 h of Mtb infection at MOI of 1 by qRT-PCR analysis. β-Actin served as an internal control. (H) Intracellular protein level was detected in Raw 264.7 at 0, 6, 24, 48, and 72 h of Mtb infection at MOI of 1 by western blot analysis. The ratio of expression of Mct4 and β-actin is shown in graph. β-Actin served as an internal control. (I) Intracellular mRNA level was detected in THP-1-Mφs at 0, 6, 24, 48, and 72 h of Mtb infection at MOI of 1 by qRT-PCR analysis. β-Actin served as an internal control. (J) Intracellular protein level was detected in THP-1-Mφs at 0, 6, 24, 48, and 72 h of Mtb infection at MOI of 1 by western blot analysis. The ratio of expression of Mct4 and β-actin is shown in graph. β-Actin served as an internal control. (K) Representative example of immunohistochemical staining for Mct4 in lymph node biopsies of nine patients with lymphatic TB and seven patients with chronic inflammation (CI). Data are quantified by area density analysis (right). Scale bars, 50 μm. (A–K) Data shown were the mean ± SD and are from at least three independent experiments. Data were analyzed by unpaired t test and ANOVA, ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001.

Journal: iScience

Article Title: Monocarboxylate transporter 4 facilitates Mycobacterium tuberculosis survival through NF-κB p65-mediated interleukin-10 production

doi: 10.1016/j.isci.2024.110238

Figure Lengend Snippet: Mct4 expression is promoted by Mtb infection and exerts potent pro-Mtb effects among SLC family in macrophages (A) 5 × 10 5 of BMDMs in 12-well plates were extracted by Trizol method to conduct the subsequent experiment. Intracellular mRNA level was detected in BMDMs at 24 h of Mtb infection at MOI of 1 by RNA sequencing. (B) 5 × 10 5 of BMDMs in 12-well plates were extracted by Trizol method to conduct the subsequent experiment. Intracellular mRNA level was detected in BMDMs at 0, 6, 24, 48, and 72 h of Mtb infection at MOI of 1 by qRT-PCR analysis. β-Actin served as an internal control. (C) SLC gene-silenced BMDMs were detected for relative mRNA expression by quantitative reverse-transcription PCR (qRT-PCR) analysis. β-Actin served as an internal control. (D) The intracellular Mtb load was detected in SLC gene-silenced BMDMs at 0 and 48 h of Mtb infection at MOI of 5 by colony-forming unit (CFU) assay. (E) Intracellular mRNA level was detected in BMDMs at 0, 6, 24, 48, and 72 h of Mtb infection at MOI of 1 by qRT-PCR analysis. β-Actin served as an internal control. (F) Intracellular protein level was detected in BMDMs at 0, 6, 24, 48, and 72 h of Mtb infection at MOI of 1 by western blot analysis. The ratio of expression of Mct4 and β-actin is shown in graph. β-Actin served as an internal control. (G) Intracellular mRNA level was detected in Raw 264.7 at 0, 6, 24, 48, and 72 h of Mtb infection at MOI of 1 by qRT-PCR analysis. β-Actin served as an internal control. (H) Intracellular protein level was detected in Raw 264.7 at 0, 6, 24, 48, and 72 h of Mtb infection at MOI of 1 by western blot analysis. The ratio of expression of Mct4 and β-actin is shown in graph. β-Actin served as an internal control. (I) Intracellular mRNA level was detected in THP-1-Mφs at 0, 6, 24, 48, and 72 h of Mtb infection at MOI of 1 by qRT-PCR analysis. β-Actin served as an internal control. (J) Intracellular protein level was detected in THP-1-Mφs at 0, 6, 24, 48, and 72 h of Mtb infection at MOI of 1 by western blot analysis. The ratio of expression of Mct4 and β-actin is shown in graph. β-Actin served as an internal control. (K) Representative example of immunohistochemical staining for Mct4 in lymph node biopsies of nine patients with lymphatic TB and seven patients with chronic inflammation (CI). Data are quantified by area density analysis (right). Scale bars, 50 μm. (A–K) Data shown were the mean ± SD and are from at least three independent experiments. Data were analyzed by unpaired t test and ANOVA, ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001.

Article Snippet: Transient small interfering RNA (siRNA) targeting mouse Slc16a3 (NM_001038653.1), human SLC16A3 (NM_001042422.3), mouse IL-10 (NM_010548.2) were synthesized by RiboBio (Guangzhou, China) according to NCBI gene database.

Techniques: Expressing, Infection, RNA Sequencing, Quantitative RT-PCR, Control, Reverse Transcription, Colony-forming Unit Assay, Western Blot, Immunohistochemical staining, Staining

Mct4 inhibits anti-Mtb effect in macrophages (A) Intracellular mRNA level was detected in Slc16a3 +/+ and Slc16a3 −/− BMDMs at 0, 6, 24, and 48 h of Mtb infection at MOI of 1 by qRT-PCR analysis. β-Actin served as an internal control. (B) Intracellular protein level was detected in Slc16a3 +/+ and Slc16a3 −/− BMDMs at 0, 6, 24, and 48 h of Mtb infection at MOI of 1 by western blot analysis. The ratio of expression of Mct4 and β-actin is shown in graph. β-Actin served as an internal control. (C) The intracellular Mtb load was detected in Slc16a3 +/+ and Slc16a3 −/− BMDMs at 0 and 48 h of Mtb infection at MOI of 5 by CFU assay. (D) Intracellular mRNA level was detected in Raw 264.7 cells overexpressing Mct4 at 0, 6, 24, and 48 h of Mtb infection at MOI of 1 by qRT-PCR analysis. β-Actin served as an internal control. (E) Intracellular protein level was detected in Raw 264.7 cells overexpressing Mct4 at 0, 6, 24, and 48 h of Mtb infection at MOI of 1 by western blot analysis. The ratio of expression of Mct4 and β-actin is shown in graph. β-Actin served as an internal control. (F) The intracellular Mtb load was detected in Raw 264.7 cells overexpressing Mct4 at 0 and 48 h of Mtb infection at MOI of 5 by CFU assay. (G) Intracellular mRNA level was detected in Mct4-silenced THP-1-Mφs at 0, 6, 24, and 48 h of Mtb infection at MOI of 1 by qRT-PCR analysis. β-Actin served as an internal control. (H) Intracellular protein level was detected in Mct4-silenced THP-1-Mφs at 0, 6, 24, and 48 h of Mtb infection at MOI of 1 by western blot analysis. The ratio of expression of Mct4 and β-actin is shown in graph. β-Actin served as an internal control. (I) The intracellular Mtb load was detected in Mct4-silenced THP-1-Mφs at 0 and 48 h of Mtb infection at MOI of 5 by CFU assay. (A–I) Data shown were the mean ± SD and are from at least three independent experiments. Data were analyzed by two-way ANOVA, ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001.

Journal: iScience

Article Title: Monocarboxylate transporter 4 facilitates Mycobacterium tuberculosis survival through NF-κB p65-mediated interleukin-10 production

doi: 10.1016/j.isci.2024.110238

Figure Lengend Snippet: Mct4 inhibits anti-Mtb effect in macrophages (A) Intracellular mRNA level was detected in Slc16a3 +/+ and Slc16a3 −/− BMDMs at 0, 6, 24, and 48 h of Mtb infection at MOI of 1 by qRT-PCR analysis. β-Actin served as an internal control. (B) Intracellular protein level was detected in Slc16a3 +/+ and Slc16a3 −/− BMDMs at 0, 6, 24, and 48 h of Mtb infection at MOI of 1 by western blot analysis. The ratio of expression of Mct4 and β-actin is shown in graph. β-Actin served as an internal control. (C) The intracellular Mtb load was detected in Slc16a3 +/+ and Slc16a3 −/− BMDMs at 0 and 48 h of Mtb infection at MOI of 5 by CFU assay. (D) Intracellular mRNA level was detected in Raw 264.7 cells overexpressing Mct4 at 0, 6, 24, and 48 h of Mtb infection at MOI of 1 by qRT-PCR analysis. β-Actin served as an internal control. (E) Intracellular protein level was detected in Raw 264.7 cells overexpressing Mct4 at 0, 6, 24, and 48 h of Mtb infection at MOI of 1 by western blot analysis. The ratio of expression of Mct4 and β-actin is shown in graph. β-Actin served as an internal control. (F) The intracellular Mtb load was detected in Raw 264.7 cells overexpressing Mct4 at 0 and 48 h of Mtb infection at MOI of 5 by CFU assay. (G) Intracellular mRNA level was detected in Mct4-silenced THP-1-Mφs at 0, 6, 24, and 48 h of Mtb infection at MOI of 1 by qRT-PCR analysis. β-Actin served as an internal control. (H) Intracellular protein level was detected in Mct4-silenced THP-1-Mφs at 0, 6, 24, and 48 h of Mtb infection at MOI of 1 by western blot analysis. The ratio of expression of Mct4 and β-actin is shown in graph. β-Actin served as an internal control. (I) The intracellular Mtb load was detected in Mct4-silenced THP-1-Mφs at 0 and 48 h of Mtb infection at MOI of 5 by CFU assay. (A–I) Data shown were the mean ± SD and are from at least three independent experiments. Data were analyzed by two-way ANOVA, ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001.

Techniques: Infection, Quantitative RT-PCR, Control, Western Blot, Expressing, Colony-forming Unit Assay

Mct4 deficiency reduced Mtb survival in mice (A) Slc16a3 +/+ and Slc16a3 −/− mice (six mice per group) aged from 6 to 8 weeks were infected with about 1 × 10 7 CFUs of the Mtb strain by aerosol, and the Mtb load of lungs and spleens at 1 week and 4 weeks post Mtb infection were detected by CFU assay. (B) Slc16a3 +/+ and Slc16a3 −/− mice (six mice per group) aged from 6 to 8 weeks were infected with about 1 × 10 7 CFUs of the Mtb strain by aerosol, and the slices of lungs and spleens at 4 weeks post Mtb infection were detected by hematoxylin-eosin staining. Multinucleated giant cells in spleens were counted in 30 different sights of slices; the count of MGCs is shown in graph. Lung: scale bars, 50 μm. Spleen: scale bars, 25 μm. (C) Surface markers (CD80, CD86, and MHC-II) of macrophages in the lungs of Slc16a3 +/+ and Slc16a3 −/− mice were analyzed by flow cytometry at 1 week upon Mtb infection. The expression of macrophage surface markers was assessed by median fluorescence intensity (MFI). Plots are gated on CD11b + F4/80 + cells. (D and E) Surface markers of (D) Slc16a3 +/+ and Slc16a3 −/− BMDMs and (E) Raw 264.7 cells overexpressing Mct4 were analyzed by flow cytometry at 24 h upon Mtb infection. (A–E) Data shown were the mean ± SD and are from at least three independent experiments. Data were analyzed by unpaired t test and two-way ANOVA, ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001.

Journal: iScience

Article Title: Monocarboxylate transporter 4 facilitates Mycobacterium tuberculosis survival through NF-κB p65-mediated interleukin-10 production

doi: 10.1016/j.isci.2024.110238

Figure Lengend Snippet: Mct4 deficiency reduced Mtb survival in mice (A) Slc16a3 +/+ and Slc16a3 −/− mice (six mice per group) aged from 6 to 8 weeks were infected with about 1 × 10 7 CFUs of the Mtb strain by aerosol, and the Mtb load of lungs and spleens at 1 week and 4 weeks post Mtb infection were detected by CFU assay. (B) Slc16a3 +/+ and Slc16a3 −/− mice (six mice per group) aged from 6 to 8 weeks were infected with about 1 × 10 7 CFUs of the Mtb strain by aerosol, and the slices of lungs and spleens at 4 weeks post Mtb infection were detected by hematoxylin-eosin staining. Multinucleated giant cells in spleens were counted in 30 different sights of slices; the count of MGCs is shown in graph. Lung: scale bars, 50 μm. Spleen: scale bars, 25 μm. (C) Surface markers (CD80, CD86, and MHC-II) of macrophages in the lungs of Slc16a3 +/+ and Slc16a3 −/− mice were analyzed by flow cytometry at 1 week upon Mtb infection. The expression of macrophage surface markers was assessed by median fluorescence intensity (MFI). Plots are gated on CD11b + F4/80 + cells. (D and E) Surface markers of (D) Slc16a3 +/+ and Slc16a3 −/− BMDMs and (E) Raw 264.7 cells overexpressing Mct4 were analyzed by flow cytometry at 24 h upon Mtb infection. (A–E) Data shown were the mean ± SD and are from at least three independent experiments. Data were analyzed by unpaired t test and two-way ANOVA, ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001.

Techniques: Infection, Aerosol, Colony-forming Unit Assay, Staining, Flow Cytometry, Expressing, Fluorescence

Mct4 promotes Mtb survival through IL-10 production in macrophages (A) Intracellular mRNA level and production of IL-1β and IL-10 were detected in Slc16a3 +/+ and Slc16a3 −/− BMDMs at 0, 6, 24, and 48 h of Mtb infection at MOI of 1 by qRT-PCR and ELISA analysis. β-Actin served as an internal control. (B) Intracellular mRNA level and production of IL-1β and IL-10 were detected in Raw 264.7 cells overexpressing Mct4 at 0, 6, 24, and 48 h of Mtb infection at MOI of 1 by qRT-PCR and ELISA analysis. Intracellular protein level was detected by western blot analysis. The ratio of expression of IL-1β and β-actin is shown in graph. β-Actin served as an internal control. (C) Intracellular mRNA level and production of IL-1β and IL-10 were detected in Slc16a3-silencing THP-1-Mφs at 0, 6, 24, and 48 h of Mtb infection at MOI of 1 by qRT-PCR and ELISA analysis. β-Actin served as an internal control. (D) Intracellular mRNA and production of IL-10 were detected in Raw 264.7 cells overexpressing Mct4 pretreated with 1 μg/mL IL-10 and isotype control antibody for 2 h upon Mtb infection at MOI of 1 for 48 h by qRT-PCR and ELISA analysis. β-Actin served as an internal control. (E) The intracellular Mtb load was detected in Raw 264.7 cells overexpressing Mct4 pretreated with 1 μg/mL IL-10 and isotype control antibody for 2 h upon Mtb infection at MOI of 5 for 48 h by CFU assay. (F) Intracellular mRNA and production of IL-10 were detected in Raw 264.7 cells overexpressing Mct4 pretreated with IL-10 siRNA for 24 h upon Mtb infection at MOI of 1 for 48 h by qRT-PCR and ELISA analysis. β-Actin served as an internal control. (G) The intracellular Mtb load was detected in Raw 264.7 cells overexpressing Mct4 pretreated with IL-10 siRNA for 24 h upon Mtb infection at MOI of 5 for 48 h by CFU assay. (A–G) Western blot results were representative of three independent experiments with similar results. Data shown were the mean ± SD and are from at least three independent experiments. Data were analyzed by two-way ANOVA, ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001; ns, not significant.

Journal: iScience

Article Title: Monocarboxylate transporter 4 facilitates Mycobacterium tuberculosis survival through NF-κB p65-mediated interleukin-10 production

doi: 10.1016/j.isci.2024.110238

Figure Lengend Snippet: Mct4 promotes Mtb survival through IL-10 production in macrophages (A) Intracellular mRNA level and production of IL-1β and IL-10 were detected in Slc16a3 +/+ and Slc16a3 −/− BMDMs at 0, 6, 24, and 48 h of Mtb infection at MOI of 1 by qRT-PCR and ELISA analysis. β-Actin served as an internal control. (B) Intracellular mRNA level and production of IL-1β and IL-10 were detected in Raw 264.7 cells overexpressing Mct4 at 0, 6, 24, and 48 h of Mtb infection at MOI of 1 by qRT-PCR and ELISA analysis. Intracellular protein level was detected by western blot analysis. The ratio of expression of IL-1β and β-actin is shown in graph. β-Actin served as an internal control. (C) Intracellular mRNA level and production of IL-1β and IL-10 were detected in Slc16a3-silencing THP-1-Mφs at 0, 6, 24, and 48 h of Mtb infection at MOI of 1 by qRT-PCR and ELISA analysis. β-Actin served as an internal control. (D) Intracellular mRNA and production of IL-10 were detected in Raw 264.7 cells overexpressing Mct4 pretreated with 1 μg/mL IL-10 and isotype control antibody for 2 h upon Mtb infection at MOI of 1 for 48 h by qRT-PCR and ELISA analysis. β-Actin served as an internal control. (E) The intracellular Mtb load was detected in Raw 264.7 cells overexpressing Mct4 pretreated with 1 μg/mL IL-10 and isotype control antibody for 2 h upon Mtb infection at MOI of 5 for 48 h by CFU assay. (F) Intracellular mRNA and production of IL-10 were detected in Raw 264.7 cells overexpressing Mct4 pretreated with IL-10 siRNA for 24 h upon Mtb infection at MOI of 1 for 48 h by qRT-PCR and ELISA analysis. β-Actin served as an internal control. (G) The intracellular Mtb load was detected in Raw 264.7 cells overexpressing Mct4 pretreated with IL-10 siRNA for 24 h upon Mtb infection at MOI of 5 for 48 h by CFU assay. (A–G) Western blot results were representative of three independent experiments with similar results. Data shown were the mean ± SD and are from at least three independent experiments. Data were analyzed by two-way ANOVA, ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001; ns, not significant.

Techniques: Infection, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Control, Western Blot, Expressing, Colony-forming Unit Assay

Mct4 enhances IL-10 production by NF-κB p65 activation in Mtb-infected macrophages (A) Intracellular protein level was detected in Slc16a3 +/+ and Slc16a3 −/− BMDMs at 0, 0.5, 1, and 2 h of Mtb infection at MOI of 5 by western blot analysis. The ratio of expression of phosphorylation of NF-κB p65 and NF-κB p65 is shown in graph. The ratio of expression of phosphorylation of p38 MAPK and p38 MAPK is shown in graph. β-Actin served as an internal control. (B) Intracellular protein level was detected in Raw 264.7 cells overexpressing Mct4 at 0, 0.5, 1, and 2 h of Mtb infection at MOI of 5 by western blot analysis. The ratio of expression of phosphorylation of NF-κB p65 and NF-κB p65 is shown in graph. The ratio of expression of phosphorylation of p38 MAPK and p38 MAPK is shown in graph. β-Actin served as an internal control. (C) Phosphorylation level of p38 MAPK was detected by western blot in Raw 264.7 cells overexpressing Mct4 pretreated with 10 μM SB203580 (p38 MAPK inhibitor) for 1 h at Mtb infection at MOI of 5 for 60 min. The ratio of expression of phosphorylation of p38 MAPK and p38 MAPK is shown in graph. β-Actin served as an internal control. (D) Phosphorylation level of NF-κB was detected by western blot in Raw 264.7 cells overexpressing Mct4 pretreated with 20 μM JSH23(NF-κB p65 inhibitor) for 1 h at Mtb infection at MOI of 5 for 60 min. The ratio of expression of phosphorylation of p38 MAPK and p38 MAPK is shown in graph. The ratio of expression of phosphorylation of NF-κB p65 and NF-κB p65 is shown in graph. β-Actin served as an internal control. (E–H) Intracellular mRNA and production of IL-10 were detected in Raw 264.7 cells overexpressing Mct4 pretreated with (E) 10 μM SB203580 and (G) 20 μM JSH23 for 1 h upon Mtb infection at MOI of 1 for 48 h by qRT-PCR and ELISA analysis. β-Actin served as an internal control. The intracellular Mtb load was detected in Raw 264.7 cells overexpressing Mct4 pretreated with (F) 10 μM SB203580 and (H) 20 μM JSH23 for 1 h upon Mtb infection at MOI of 5 for 48 h by CFU assay. (I) Slc16a3 +/+ and Slc16a3 −/− mice (six mice per group) aged from 6 to 8 weeks were infected with about 1 × 10 7 CFUs of the Mtb strain by aerosol and were analyzed 4 weeks later. Left: F4/80 and phosphorylation of NF-κB p65 protein in the lungs of Slc16a3 +/+ and Slc16a3 −/− mice was assessed by dual immunofluorescence technique. Right: dual immunofluorescence maps were quantified by the Pearson’s correlation coefficient. At least five visual fields were randomly from each slide in each group for analysis. Scale bars, 50 μm. (A–I) Western blot results were representative of three independent experiments with similar results. Data shown were the mean ± SD and are from at least three independent experiments. Data were analyzed by unpaired t test and two-way ANOVA, ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001; ns, not significant.

Journal: iScience

Article Title: Monocarboxylate transporter 4 facilitates Mycobacterium tuberculosis survival through NF-κB p65-mediated interleukin-10 production

doi: 10.1016/j.isci.2024.110238

Figure Lengend Snippet: Mct4 enhances IL-10 production by NF-κB p65 activation in Mtb-infected macrophages (A) Intracellular protein level was detected in Slc16a3 +/+ and Slc16a3 −/− BMDMs at 0, 0.5, 1, and 2 h of Mtb infection at MOI of 5 by western blot analysis. The ratio of expression of phosphorylation of NF-κB p65 and NF-κB p65 is shown in graph. The ratio of expression of phosphorylation of p38 MAPK and p38 MAPK is shown in graph. β-Actin served as an internal control. (B) Intracellular protein level was detected in Raw 264.7 cells overexpressing Mct4 at 0, 0.5, 1, and 2 h of Mtb infection at MOI of 5 by western blot analysis. The ratio of expression of phosphorylation of NF-κB p65 and NF-κB p65 is shown in graph. The ratio of expression of phosphorylation of p38 MAPK and p38 MAPK is shown in graph. β-Actin served as an internal control. (C) Phosphorylation level of p38 MAPK was detected by western blot in Raw 264.7 cells overexpressing Mct4 pretreated with 10 μM SB203580 (p38 MAPK inhibitor) for 1 h at Mtb infection at MOI of 5 for 60 min. The ratio of expression of phosphorylation of p38 MAPK and p38 MAPK is shown in graph. β-Actin served as an internal control. (D) Phosphorylation level of NF-κB was detected by western blot in Raw 264.7 cells overexpressing Mct4 pretreated with 20 μM JSH23(NF-κB p65 inhibitor) for 1 h at Mtb infection at MOI of 5 for 60 min. The ratio of expression of phosphorylation of p38 MAPK and p38 MAPK is shown in graph. The ratio of expression of phosphorylation of NF-κB p65 and NF-κB p65 is shown in graph. β-Actin served as an internal control. (E–H) Intracellular mRNA and production of IL-10 were detected in Raw 264.7 cells overexpressing Mct4 pretreated with (E) 10 μM SB203580 and (G) 20 μM JSH23 for 1 h upon Mtb infection at MOI of 1 for 48 h by qRT-PCR and ELISA analysis. β-Actin served as an internal control. The intracellular Mtb load was detected in Raw 264.7 cells overexpressing Mct4 pretreated with (F) 10 μM SB203580 and (H) 20 μM JSH23 for 1 h upon Mtb infection at MOI of 5 for 48 h by CFU assay. (I) Slc16a3 +/+ and Slc16a3 −/− mice (six mice per group) aged from 6 to 8 weeks were infected with about 1 × 10 7 CFUs of the Mtb strain by aerosol and were analyzed 4 weeks later. Left: F4/80 and phosphorylation of NF-κB p65 protein in the lungs of Slc16a3 +/+ and Slc16a3 −/− mice was assessed by dual immunofluorescence technique. Right: dual immunofluorescence maps were quantified by the Pearson’s correlation coefficient. At least five visual fields were randomly from each slide in each group for analysis. Scale bars, 50 μm. (A–I) Western blot results were representative of three independent experiments with similar results. Data shown were the mean ± SD and are from at least three independent experiments. Data were analyzed by unpaired t test and two-way ANOVA, ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001; ns, not significant.

Techniques: Activation Assay, Infection, Western Blot, Expressing, Phospho-proteomics, Control, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Colony-forming Unit Assay, Aerosol, Immunofluorescence

Mct4 activates p65-IL-10 axis to promote Mtb survival by translocating intracellular lactate out of macrophages (A) Intracellular and extracellular lactate level was detected in Slc16a3 +/+ and Slc16a3 −/− BMDMs at 0, 24, and 48 h of Mtb infection at MOI of 1 by lactate assay kit. (B) Intracellular and extracellular lactate level was detected in Raw 264.7 cells overexpressing Mct4 at 0, 24, and 48 h of Mtb infection at MOI of 1 by lactate assay kit. (C) Phosphorylation level of NF-κB p65 was detected by western blot in Raw 264.7 cells overexpressing Mct4 pretreated with 25 mM lactate for 3 h upon Mtb infection at MOI of 5 for 60 min. The ratio of expression of phosphorylation of NF-κB p65 and NF-κB p65 is shown in graph. β-Actin served as an internal control. (D) Intracellular mRNA and production of IL-10 were detected in Raw 264.7 cells overexpressing Mct4 pretreated with 25 mM lactate for 3 h upon Mtb infection at MOI of 1 for 48 h by qRT-PCR and ELISA analysis. β-Actin served as an internal control. (E) The intracellular Mtb load was detected in Raw 264.7 cells overexpressing Mct4 pretreated with 25 mM lactate for 3 h upon Mtb infection at MOI of 5 for 48 h by CFU assay. (A–E) Western blot results were representative of three independent experiments with similar results. Data shown were the mean ± SD and are from at least three independent experiments. Data were analyzed by unpaired t test and two-way ANOVA, ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001; ns, not significant.

Journal: iScience

Article Title: Monocarboxylate transporter 4 facilitates Mycobacterium tuberculosis survival through NF-κB p65-mediated interleukin-10 production

doi: 10.1016/j.isci.2024.110238

Figure Lengend Snippet: Mct4 activates p65-IL-10 axis to promote Mtb survival by translocating intracellular lactate out of macrophages (A) Intracellular and extracellular lactate level was detected in Slc16a3 +/+ and Slc16a3 −/− BMDMs at 0, 24, and 48 h of Mtb infection at MOI of 1 by lactate assay kit. (B) Intracellular and extracellular lactate level was detected in Raw 264.7 cells overexpressing Mct4 at 0, 24, and 48 h of Mtb infection at MOI of 1 by lactate assay kit. (C) Phosphorylation level of NF-κB p65 was detected by western blot in Raw 264.7 cells overexpressing Mct4 pretreated with 25 mM lactate for 3 h upon Mtb infection at MOI of 5 for 60 min. The ratio of expression of phosphorylation of NF-κB p65 and NF-κB p65 is shown in graph. β-Actin served as an internal control. (D) Intracellular mRNA and production of IL-10 were detected in Raw 264.7 cells overexpressing Mct4 pretreated with 25 mM lactate for 3 h upon Mtb infection at MOI of 1 for 48 h by qRT-PCR and ELISA analysis. β-Actin served as an internal control. (E) The intracellular Mtb load was detected in Raw 264.7 cells overexpressing Mct4 pretreated with 25 mM lactate for 3 h upon Mtb infection at MOI of 5 for 48 h by CFU assay. (A–E) Western blot results were representative of three independent experiments with similar results. Data shown were the mean ± SD and are from at least three independent experiments. Data were analyzed by unpaired t test and two-way ANOVA, ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001; ns, not significant.

Techniques: Infection, Lactate Assay, Phospho-proteomics, Western Blot, Expressing, Control, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Colony-forming Unit Assay

Potential mechanism for Mct4 to promote Mtb survival in macrophages Mtb infection increases Mct4 expression in macrophages, and Mct4 transports lactate outside of macrophages to decrease intracellular lactate, which promotes NF-κB p65 activation and IL-10 production, eventually facilitating Mtb survival.

Journal: iScience

Article Title: Monocarboxylate transporter 4 facilitates Mycobacterium tuberculosis survival through NF-κB p65-mediated interleukin-10 production

doi: 10.1016/j.isci.2024.110238

Figure Lengend Snippet: Potential mechanism for Mct4 to promote Mtb survival in macrophages Mtb infection increases Mct4 expression in macrophages, and Mct4 transports lactate outside of macrophages to decrease intracellular lactate, which promotes NF-κB p65 activation and IL-10 production, eventually facilitating Mtb survival.

Techniques: Infection, Expressing, Activation Assay

Journal: iScience

Article Title: Monocarboxylate transporter 4 facilitates Mycobacterium tuberculosis survival through NF-κB p65-mediated interleukin-10 production

doi: 10.1016/j.isci.2024.110238

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