Journal: Scientific Reports

Article Title: Cholangiocarcinoma derived exosomes attenuate the anti-tumor functions of NK cells

doi: 10.1038/s41598-026-36706-9

Figure Lengend Snippet: Effect of RBE-Exos on the adhesion of NK-92 cells. ( A ) Growth status of NK-92 cells. NK cells treated with RBE-Exos lost the normal cluster growing. ( B ) CD11a protein levels. ( C ) CD18 protein levels. ( D ) CD54 protein levels. Western blot showed that the protein levels of CD11a, CD18 and, CD54 were obviously decreased in NK-92 cells treated with RBE-Exos. ( E ) Relative transcriptional levels of CD11a , CD18 , CD54 and, CD2 in NK-92 cells. qRT-PCR displayed that RBE-Exos downregulated the transcriptional levels of CD11a , CD18 , CD54 , but not CD2 . Data were presented as mean ± SEM for three independent experiments. * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: The primary antibodies were as follows: rabbit anti-human CD63, BCL-2, BCL-XL, CD11a, β-actin (Abcam, USA), mouse anti-human HSP70 (Santa-Cruz, USA), rabbit anti-human CD18 (Proteintech, China), mouse anti-human CD54 (Proteintech, China).

Techniques: Western Blot, Quantitative RT-PCR

Journal: Scientific Reports

Article Title: Cholangiocarcinoma derived exosomes attenuate the anti-tumor functions of NK cells

doi: 10.1038/s41598-026-36706-9

Figure Lengend Snippet: Cytotoxicity and expression of adhesion molecules in primary NK cells. ( A ) Cytotoxicity of primary NK cells from CCA patients and healthy people against target cells. LDH detection displayed the cytotoxicity of NK cells from CCA patients was significantly lower than that of NK cells healthy people. Data are presented as the mean ± SEM from n = 3 independent experiments. ( B ) Protein levels of CD18 in primary NK cells from CCA patients and healthy people. Data are presented as the mean ± SEM from n = 3 independent experiments. ( C ) Protein levels of CD54 in primary NK cells from CCA patients and healthy people. Data are presented as the mean ± SEM from n = 3 independent experiments. ** P < 0.01.

Article Snippet: The primary antibodies were as follows: rabbit anti-human CD63, BCL-2, BCL-XL, CD11a, β-actin (Abcam, USA), mouse anti-human HSP70 (Santa-Cruz, USA), rabbit anti-human CD18 (Proteintech, China), mouse anti-human CD54 (Proteintech, China).

Techniques: Expressing

Journal: International Journal of Ophthalmology

Article Title: Pirfenidone suppresses the abnormal activation of human Müller cells after platelet-derived growth factor-BB stimulation

doi: 10.18240/ijo.2019.07.04

Figure Lengend Snippet: A: The cells of the normal group and the model group were respectively treated by 0, 0.2 and 0.3 mg/mL pirfenidone. The relative mRNA levels of TGF-β1, TGF-β2 and PEDF were determined by real-time PCR. Results are shown as indicated. The expression levels of TGF-β1, TGF-β2 and PEDF under the indicated treatment of pirfenidone were detected by Western blotting (B) and immunofluorescence (C). Green fluorescence, for the indicated proteins; blue fluorescence for the nucleus. aP<0.05 vs normal+0 mg/mL pirfenidone group, bP<0.01 vs normal+0 mg/mL pirfenidone group, cP<0.0001 vs normal+0 mg/mL pirfenidone group; dP<0.05 vs model+0 mg/mL pirfenidone group, eP<0.01 vs model+0 mg/mL pirfenidone group, fP<0.0001 vs model+0 mg/mL pirfenidone group; gP<0.05 vs model+0.2 mg/mL pirfenidone group, hP<0.01 vs model+0.2 mg/mL pirfenidone group, iP<0.0001 vs model+0.2 mg/mL pirfenidone group.

Article Snippet: The expression of TGF-β1, TGF-β2, PEDF, and β-actin was respectively tested by anti-TGF-β1 antibody (Boster, USA, {"type":"entrez-nucleotide","attrs":{"text":"A04630","term_id":"411029","term_text":"A04630"}} A04630 ), anti-TGF-β2 antibody (Boster, USA, {"type":"entrez-protein","attrs":{"text":"A00892","term_id":"77211","term_text":"pir||A00892"}} A00892 ), anti-PEDF antibody (Abcam, USA, ab233120) and anti-β-actin antibody (Invitrogen, USA, PA1183).

Techniques: Real-time Polymerase Chain Reaction, Expressing, Western Blot, Immunofluorescence, Fluorescence

Journal: Frontiers in Physiology

Article Title: Atlas of human dental pulp cells at multiple spatial and temporal levels based on single-cell sequencing analysis

doi: 10.3389/fphys.2022.993478

Figure Lengend Snippet: The top 20 genes in odontoblasts.

Article Snippet: The following antibodies were used in our study: CD163 (1:500, Abcam, United States), SLC12A2 (1:100, Proteintech, China), ST8SIA1 (1:100, Proteintech), CD24 (1: 50, Santa Cruz, United States), WISP1 (1:100, Proteintech), CD146/MCAM (1: 250, Abcam), and CD90 (1:200, Abcam).

Techniques:

Journal: Frontiers in Genetics

Article Title: Cell-based analysis of CLIC5A and SLC12A2 variants associated with hearing impairment in two African families

doi: 10.3389/fgene.2022.924904

Figure Lengend Snippet: Mutant (MT) CLIC5A and SLC12A2 proteins are expressed at higher levels in HEK-293 cells, relative to the wild type proteins (WT). (A) Western blot of total proteins from HEK-293 cells transfected with wild-type (WT) or (C) 224T>C; p.(L75P) mutant (MT) CLIC5A plasmids; using anti-Myc antibody (N-terminal). (B) Densitometric analysis of western blot of CLIC5. (C) Western blot of total protein isolated from HEK-293 cells transfected with WT and (C) 2935G>A:p.(E979K) MT SLC12A2 plasmids; using anti-flag antibody (N-terminal). (D) Densitometric analysis of western blots of SLC12A2 for the WT and MT SLC12A2 treated cells using ImageJ. The uncropped western blot pictures are shown in . Schematic diagrams showing (E) CLIC5 p.(L75P) and (F) SLC12A2 p(E979K) variant positions. The protein domains were predicted using the Protein Families Database (Pfam) ( https://doi.org/10.1093/nar/gkaa913 ).

Article Snippet: Mammalian expression constructs containing human CLIC5A (NM_001256023, UniProtKB - Q9NZA1) and SLC12A2 (NM_001046, UniProtKB - P55011) cDNAs were purchased from ORIGENE (Rockville, Maryland).

Techniques: Mutagenesis, Western Blot, Transfection, Isolation, Variant Assay

Journal: Frontiers in Genetics

Article Title: Cell-based analysis of CLIC5A and SLC12A2 variants associated with hearing impairment in two African families

doi: 10.3389/fgene.2022.924904

Figure Lengend Snippet: SLC12A2 WT HEK293 expressing cells display distinct morphology not observed in mutant-expressing cells. Live HEK293 cells expressing WT or MT GFP-tagged SLC12A2 protein and stained with Hoechst were observed using confocal microscopy 72 h post transfection. The red arrows point to axon-like structures present in the WT- SLC12A2 population of cells.

Article Snippet: Mammalian expression constructs containing human CLIC5A (NM_001256023, UniProtKB - Q9NZA1) and SLC12A2 (NM_001046, UniProtKB - P55011) cDNAs were purchased from ORIGENE (Rockville, Maryland).

Techniques: Expressing, Mutagenesis, Staining, Confocal Microscopy, Transfection

Journal: Frontiers in Genetics

Article Title: Cell-based analysis of CLIC5A and SLC12A2 variants associated with hearing impairment in two African families

doi: 10.3389/fgene.2022.924904

Figure Lengend Snippet: Mutant (MT) SLC12A2 expressing cells had relatively less phosphorylated p38 (Pp38) compared to the wild type (WT). (A) Western blot showing Pp38 expression in HEK293 cells transfected with WT and (C) 2935G>A: p.(E979K) MT SLC12A2 plasmids. Total p38 was used for normalization (B) Densitometric analysis of western blots of Pp38 the WT and MT SLC12A2 treated cells using ImageJ. The densitometric analysis was conducted 3 times and the mean measurements were recorded. The uncropped western blot pictures are shown in .

Article Snippet: Mammalian expression constructs containing human CLIC5A (NM_001256023, UniProtKB - Q9NZA1) and SLC12A2 (NM_001046, UniProtKB - P55011) cDNAs were purchased from ORIGENE (Rockville, Maryland).

Techniques: Mutagenesis, Expressing, Western Blot, Transfection

Journal: Frontiers in Genetics

Article Title: Cell-based analysis of CLIC5A and SLC12A2 variants associated with hearing impairment in two African families

doi: 10.3389/fgene.2022.924904

Figure Lengend Snippet: Association of CLIC5 and SLC12A2 variants with hearing impairment in patients.

Article Snippet: Mammalian expression constructs containing human CLIC5A (NM_001256023, UniProtKB - Q9NZA1) and SLC12A2 (NM_001046, UniProtKB - P55011) cDNAs were purchased from ORIGENE (Rockville, Maryland).

Techniques: