Journal: Inflammation

Article Title: HNRNPC-Mediated m6A Epitranscriptomics Drives CD80-Dependent Tubular Dysfunction in Sepsis-Induced AKI

doi: 10.1007/s10753-025-02349-7

Figure Lengend Snippet: HNRNPC induces apoptosis via targeting CD80. ( A ) Heatmap showing the cluster distribution. ( B ) Volcano plot showing the differentially expressed genes between HK2 cells with/without HNRNPC knockdown. ( C and D ) KEGG ( C ) and GO ( D ) functional enrichment analyses. ( E – G ) CD80 was upregulated in AKI mice, as measured by IHC ( E ), WB ( F ) and ELISA ( G ). ( H and I ) CD80 was up-regulated in HK2 cells treating with LPS, and was then knocked down, as measured by q-PCR ( H ) and WB ( I ). ( J ) Cell viability of sh-CD80/sh-NC-expressing HK2 cells following induction with or without LPS. ( K and L ) Apoptosis-related protein levels ( K ) and F-actin protein level ( L ) in HK2 cells with or without CD80 knockdown following treating with LPS, as measured by WB. ( M ) Cell viability of sh-HNRNPC-expressing HK2 cells with CD80 over-expression following induction with of LPS. ( N and O ) Apoptosis-related protein levels ( N ) and F-actin protein level ( O ) in sh-HNRNPC-expressing HK2 cells with CD80 over-expression following induction with of LPS. Data are presented as means from three independent experiments. Each point represented an independent data. * P < 0.05, ** P < 0.01, and *** P < 0.001

Article Snippet: Enzyme-Linked Immunosorbent Assay (ELISA) Mouse CD80 levels (CUSABIO, #CSB-EL004959MO), IL-6 levels (Thermo Fisher Scientific, #KMC0061), TNF-α levels (Thermo Fisher Scientific, #BMS607-3), and Scr levels (Jiancheng, #C011-2–1) in mice were measured following the provided instructions.

Techniques: Knockdown, Functional Assay, Enzyme-linked Immunosorbent Assay, Expressing, Over Expression

Journal: Inflammation

Article Title: HNRNPC-Mediated m6A Epitranscriptomics Drives CD80-Dependent Tubular Dysfunction in Sepsis-Induced AKI

doi: 10.1007/s10753-025-02349-7

Figure Lengend Snippet: HNRNPC promotes CD80 at transcriptional level. ( A and B ) CD80 mRNA levels in sh-HNRNPC/sh-NC-expressing HK2 cells following treatment with or without LPS, as measured by qPCR ( A ) and WB ( B ). ( C ) CD80 promoter activities in HK2 cells following the treatment with or without LPS, as measured by luciferase assays. ( D ) CD80 promoter activities in HK2 cells in the absence or presence of α-Amanitin, as measured by luciferase assays. ( E and F ) HNRNPC did not affect the protein stability of CD80. HK2 cells were treated with CHX (100 μM) during administration with or without LPS. The ratio between CD80 and GAPDH was graphed in the middle panel, and the relative CD80 level with or without CHX was also calculated in HK2 cells treated with or without LPS (right panel). Data are presented as means from three independent experiments. Each point represented an independent data. ns P ≥ 0.05 and *** P < 0.001

Article Snippet: Enzyme-Linked Immunosorbent Assay (ELISA) Mouse CD80 levels (CUSABIO, #CSB-EL004959MO), IL-6 levels (Thermo Fisher Scientific, #KMC0061), TNF-α levels (Thermo Fisher Scientific, #BMS607-3), and Scr levels (Jiancheng, #C011-2–1) in mice were measured following the provided instructions.

Techniques: Expressing, Luciferase

Journal: Inflammation

Article Title: HNRNPC-Mediated m6A Epitranscriptomics Drives CD80-Dependent Tubular Dysfunction in Sepsis-Induced AKI

doi: 10.1007/s10753-025-02349-7

Figure Lengend Snippet: HNRNPC promotes CD80 transcription via NF-κB. ( A ) The specific binding site bases of transcription factor NF-κB p65, downloaded from the JASPAR website ( https://jaspar.genereg.net/ ). ( B ) CD80 promoter activities with or without intact NF-κB p65 motif with NF-κB p65 overexpressed or knocked down in HK2 cells, as measured by luciferase assay. ( C and D ) NF-κB p65 ( C ) and CD80 ( D ) mRNA levels with NF-κB p65 overexpressed or knocked down in HK2 cells, as measured by qPCR. ( E ) NF-κB p65 and CD80 protein levels with NF-κB p65 overexpressed or knocked down in HK2 cells, as measured by WB. ( F ) The interaction between probes compassing the NF-κB p65 motif and nuclear exacts (NE) from HK2 cells, as measured by EMSA. The proportion of wild-type (Wt) probes and mutant (mut) probes in each assay was different. ( G and H ) ChIP assays using anti-NF-κB p65 antibody and control IgG antibody in HK2 cells with or without LPS ( G ). qPCR was used to quantify immunoprecipitated chromatin fragments using primers specific to NF-κB p65 binding sites ( H ). ( I - K ) HNRNPC promotes CD80 promoter activities and expression via NF-κB in HK2 cells. CD80 promoter activities, mRNA levels, and protein levels in sh-p65/sh-NC-expressing HK cells with or without inducible HNRNPC expression, as measured by luciferase assay ( I ), qPCR ( J ), and WB ( K ) respectively. Data are presented as means from three independent experiments. Each point represented an independent data. ns P ≥ 0.05 and *** P < 0.001

Article Snippet: Enzyme-Linked Immunosorbent Assay (ELISA) Mouse CD80 levels (CUSABIO, #CSB-EL004959MO), IL-6 levels (Thermo Fisher Scientific, #KMC0061), TNF-α levels (Thermo Fisher Scientific, #BMS607-3), and Scr levels (Jiancheng, #C011-2–1) in mice were measured following the provided instructions.

Techniques: Binding Assay, Luciferase, Mutagenesis, Control, Immunoprecipitation, Expressing

Journal: Inflammation

Article Title: HNRNPC-Mediated m6A Epitranscriptomics Drives CD80-Dependent Tubular Dysfunction in Sepsis-Induced AKI

doi: 10.1007/s10753-025-02349-7

Figure Lengend Snippet: HNRNPC facilitates apoptosis through m6A-dependent regulation of NF-κB p65 mRNA. ( A and B ) NF-κB p56 protein expression in the CLP and Sham mice, as measured by IHC ( A ; Scale bar, 100 μm) and WB ( B ). ( C and D ) The mRNA and protein levels of NF-κB p56 and CD80 were measured using qPCR ( C ) and WB ( D ) in HK2 cells with HNRNPC knocked down and treated with or without DAA for 12 h. ( E ) Cell viabilities of sh-METTL3/sh-NC-expressing HK2 cells with the treatment of LPS in the absence or presence of Z-VAD-FMK, as measured by CCK-8 assay. ( F – H ) Apoptosis levels ( F and G ) and F-actin protein level ( H ) in sh-METTL3/sh-NC-expressing HK2 cells with or without the indicated concentration of LPS in the absence or presence of Z-VAD-FMK. Data are presented as means from three independent experiments. Each point represented an independent data. ns P ≥ 0.05, ** P < 0.01, and *** P < 0.001

Article Snippet: Enzyme-Linked Immunosorbent Assay (ELISA) Mouse CD80 levels (CUSABIO, #CSB-EL004959MO), IL-6 levels (Thermo Fisher Scientific, #KMC0061), TNF-α levels (Thermo Fisher Scientific, #BMS607-3), and Scr levels (Jiancheng, #C011-2–1) in mice were measured following the provided instructions.

Techniques: Expressing, CCK-8 Assay, Concentration Assay

Journal: Inflammation

Article Title: HNRNPC-Mediated m6A Epitranscriptomics Drives CD80-Dependent Tubular Dysfunction in Sepsis-Induced AKI

doi: 10.1007/s10753-025-02349-7

Figure Lengend Snippet: Inhibiting HNRNPC expression is a potential strategy to protect against renal septic injury in vivo . ( A ) Representative images of H&E and PAS staining of kidney tissues from mice injected with Lv-HNRNPC or Lv-NC. Scale bar, 50 μm. ( B ) Expression of HNRNPC in the CLP kidney injected with Lv-HNRNPC or Lv-NC as control, as measured by WB. ( C ) Renal function serum creatinine (Scr) level in mice injected with Lv-HNRNPC or Lv-NC. ( D ) The expression level of NF-κB p65 mRNA in kidney tissues from mice injected with Lv-HNRNPC or Lv-NC. ( E and F ) CD80 mRNA ( E ) and serum CD80 ( F ) in mice injected with Lv-HNRNPC or Lv-NC. ( G and H ) IL-6 ( G ), and TNF-α ( H ) mRNA levels in kidney tissues from mice injected with Lv-HNRNPC or Lv-NC. ( I and J ) The serum IL-6 ( I ), and TNF-α ( J ) levels in mice injected with Lv-HNRNPC or Lv-NC. Data are presented as means from three independent experiments. Each point represented an independent data. *** P < 0.001

Article Snippet: Enzyme-Linked Immunosorbent Assay (ELISA) Mouse CD80 levels (CUSABIO, #CSB-EL004959MO), IL-6 levels (Thermo Fisher Scientific, #KMC0061), TNF-α levels (Thermo Fisher Scientific, #BMS607-3), and Scr levels (Jiancheng, #C011-2–1) in mice were measured following the provided instructions.

Techniques: Expressing, In Vivo, Staining, Injection, Control

Journal: Journal of Clinical Medicine

Article Title: Validation of a Ready-to-Use Lyophilized Kit for Labeling IL2 with 68 Ga: A New Avenue for Imaging Activated T-lymphocytes in Tumor Microenvironment

doi: 10.3390/jcm14165658

Figure Lengend Snippet: Flow cytometric analysis of Treg purity and CD25 expression. The cytograms show, in the upper line, CD4 vs. dump channel markers (dead cell dye, CD14/CD16/CD19/CD8), and, in the lower line, CD25 vs. FOXP3 expression in gated CD4 T-cells. Right, histogram overlay of CD25 expression in the indicated cell subsets; numbers shown the geometric mean fluorescence intensity.

Article Snippet: - CD8+ T-lymphocytes were isolated using the CD8+ T Cell Isolation Kit, human 130-096-495 for 10 9 total cells (Miltenyi Biotec).

Techniques: Expressing, Fluorescence