Journal: Scientific Reports

Article Title: Combining DNMT and HDAC6 inhibitors increases anti-tumor immune signaling and decreases tumor burden in ovarian cancer

doi: 10.1038/s41598-020-60409-4

Figure Lengend Snippet: DNMT1 protein levels are decreased by combination treatment of DNMTi and HDAC6i. ( A ) Ovarian cancer cell lines were treated as in Fig. and protein was extracted at Day 7 after treatment with IFN-gamma (IFN-γ+) (to assess MHC I and PD-L1 expression, in later figures) or control (IFN-γ -). Protein was isolated and immunoblots were run for the DNMT1 protein and α-tubulin as a loading control. Immunoblot membranes were cut and probed separately for DNMT1 (about 188 kDa) and α-tubulin (50 kDa). Cropped blots are shown here, and black lines indicate where one part of the blot ends and another begins. Figure shows the entire blot images. ( B ) The TykNu cell line was treated as in ( A ) and the protein synthesis cycloheximide added to cells on Day 7 for 0, 4, and 8 hours at 10 μM as indicated on the blot. Protein was isolated and immunoblots were run for the DNMT1 protein and α-tubulin as a loading control. Immunoblot membranes were cut and probed separately for DNMT1 (about 188 kDa) and α-tubulin (50 kDa). Cropped blots are shown here, and black lines indicate where one part of the blot ends and another begins. Figure shows the entire blot images. ( C ) Stable knockdowns of the HDAC6 protein were generated in the ID8 Trp53+/+ and Trp53−/− cell lines . Protein was extracted and immunoblots were run for the DNMT1 protein with B-actin as a loading control. Immunoblot membranes were probed for DNMT1 (about 188 kDa) and α-tubulin (50 kDa). Cropped blots are shown here and black lines indicate where one part of the blot ends and another begins. Figure shows the entire blot images. ( D ) Immunoblot showing knockdown of HDAC6 protein with a-Tubulin as a loading control. Protein was extracted and immunoblots were run for the HDAC6 protein with B-actin as a loading control. Immunoblots were probed for HDAC6 (131 kDa) and tubulin (50 kDa). Cropped blots are shown here and black lines indicate where one part of the blot ends and another begins. Figure shows the entire blot images. ( E ) Ovarian cancer cell lines were treated as in Fig. and RNA was extracted at Day 7. qRT-PCR was run for DNMT1, DNMT3a, and DNMT3b and TBP was used as a reference gene. *p < 0.05 compared to Mock.

Article Snippet: The antibodies used for immunoblotting included: DNMT1 (Sigma, D4692), PD-L1 (ProSci, 4059), HDAC1 (Cell Signaling, 2062), HDAC2 (Cell Signaling, 2540), HDAC6 (Assay Biotech, C0026), acetyl-alpha Tubulin (Cell Signaling, 3971), alpha-Tubulin (Cell Signaling, 3873).

Techniques: Expressing, Control, Isolation, Western Blot, Generated, Knockdown, Quantitative RT-PCR

Journal: Oncotarget

Article Title: Circulating programmed death ligand-1 (cPD-L1) in non-small-cell lung cancer (NSCLC)

doi: 10.18632/oncotarget.24785

Figure Lengend Snippet: Baseline PD-L1 plasma levels according to the number of metastatic sites

Article Snippet: The level of PD-L1 was measured in plasma samples using the human (PD-L1/CD274) ELISA kit (CUSABIO, MD, USA).

Techniques:

Journal: Oncotarget

Article Title: Circulating programmed death ligand-1 (cPD-L1) in non-small-cell lung cancer (NSCLC)

doi: 10.18632/oncotarget.24785

Figure Lengend Snippet: PD-L1 plasma levels at baseline ( A ) and after 3 months of first-line treatment ( B ) in subgroup of patients receiving chemotherapy (red) or no chemotherapy (blue) agents.

Article Snippet: The level of PD-L1 was measured in plasma samples using the human (PD-L1/CD274) ELISA kit (CUSABIO, MD, USA).

Techniques:

Journal: Clinical and Experimental Immunology

Article Title: CMTM6: increased circulating level and up-regulated expression in labial salivary glands in patients with primary Sjogren’s syndrome

doi: 10.1093/cei/uxab003

Figure Lengend Snippet: Expression level of serum CMTM6, PD-L1, and PD-1 in pSS patients, non-pSS and HC. ( A ) Serum CMTM6 levels were significantly higher in pSS patients (257.04 [123.22, 704.41] pg/ml) compared to those in HC (65.02 [29.59, 121.35] pg/ml). ( B ) Serum PD-L1 levels in HC (86.51 ± 29.26 pg/ml) were also decreased than those in pSS patients (248.94 ± 114.73 pg/ml). ( C ) Serum PD-1 levels in HC (64.35 ± 26.33 pg/ml) were lower than those in pSS patients (93.72 [58.83, 153.62] pg/ml). ( D–E ) Correlation between CMTM6 and PD-L1 level ( r = 0.402, P = 0.004) and correlation between PD-1 and PD-L1 level ( r = 0.337, P = 0.017). Results are presented as the mean (SD) and median (IQR). Mann–Whitney U -test and Student t -test; Spearman Correlation. ∗∗ P < 0.01, ∗∗∗ P < 0.001, ns: not statistical significant.

Article Snippet: CMTM6 antibody (NBP1-31183) and CMTM6 human ELISA kit (NBP2-75298) were purchased from NOVUS; PD-1 antibody (66220-1-Ig), PD-L1 antibody (66248-1-Ig), CD8 antibody (66868-1-AP), CD20 antibody (24828-1-AP), and PD-L1 human ELISA kit (KE00074) were commercially from Proteintech; CD4 antibodies were purchased from Immunolway (YT0762); PD-1 human ELISA kit was bought from Invitrogen (BMS2214).

Techniques: Expressing, MANN-WHITNEY

Journal: Clinical and Experimental Immunology

Article Title: CMTM6: increased circulating level and up-regulated expression in labial salivary glands in patients with primary Sjogren’s syndrome

doi: 10.1093/cei/uxab003

Figure Lengend Snippet: Immunohistochemical staining(brown) of B, T Cells marker, CD20 ( A ), CD4 ( B ), and CD8 ( C ) in infiltrating cells and CMTM6 ( D ), PD-L1 ( E ), PD-1 ( F ) expression in minor salivary gland pSS patients (right) and non-pSS patients (left). Boxed areas represented the sites of the zoomed -in images in the right. All sections are shown at 200× (left) and 400× (right) magnifications. (A–C) The expressions of CD20, CD4, and CD8 in the minor salivary glands of pSS patients and non-pSS controls. (D) The expressions of CMTM6 tended to highly expressed on dual cells both in pSS and non-pSS and CMTM6 showed much stronger expression in ductal epithelial cells around periphery of lymphocytic foci of pSS patients. (E) The PD-L1 preotein expressed both in epithelia cells and periphery of lymphocytic foci as well as had higher expression in the glands of pSS patients. (F) The PD-1 strongly expressed in lymphocytic foci and showed higher expression in the glands of pSS. The PD-1-positve staining cells also located in the sites with CD20, CD4, and CD8-positive staining sites.

Article Snippet: CMTM6 antibody (NBP1-31183) and CMTM6 human ELISA kit (NBP2-75298) were purchased from NOVUS; PD-1 antibody (66220-1-Ig), PD-L1 antibody (66248-1-Ig), CD8 antibody (66868-1-AP), CD20 antibody (24828-1-AP), and PD-L1 human ELISA kit (KE00074) were commercially from Proteintech; CD4 antibodies were purchased from Immunolway (YT0762); PD-1 human ELISA kit was bought from Invitrogen (BMS2214).

Techniques: Immunohistochemical staining, Staining, Marker, Expressing

Journal: Clinical and Experimental Immunology

Article Title: CMTM6: increased circulating level and up-regulated expression in labial salivary glands in patients with primary Sjogren’s syndrome

doi: 10.1093/cei/uxab003

Figure Lengend Snippet: CMTM6, PD-L1, and PD-1 expression in labial gland of pSS patients with different CM grades and ESSDAI scores. ( A ) CMTM6 expression in CM = 4 ( n = 29) group was significantly higher than those in CM<2 ( n = 9) and CM = 3 ( n = 12) groups. ( B ) PD-L1 expression in CM = 4 group was higher than in those in CM<2 and CM = 3 groups but there was no expressional difference between GM<2 and GM = 3 groups. ( C ) PD-1 expression show significance between CM = 4 group and CM = 3 group but there was no difference between CM<2 and CM = 3 groups. ( D ) CMTM6 expression with ESSDAI scores>14 showed significance compared to those with ESSDAI scores>14 or 514 and those with 514 showed higher PD-1 expression than those patients with 5

Article Snippet: CMTM6 antibody (NBP1-31183) and CMTM6 human ELISA kit (NBP2-75298) were purchased from NOVUS; PD-1 antibody (66220-1-Ig), PD-L1 antibody (66248-1-Ig), CD8 antibody (66868-1-AP), CD20 antibody (24828-1-AP), and PD-L1 human ELISA kit (KE00074) were commercially from Proteintech; CD4 antibodies were purchased from Immunolway (YT0762); PD-1 human ELISA kit was bought from Invitrogen (BMS2214).

Techniques: Expressing, MANN-WHITNEY