Journal: Cell death & disease

Article Title: The novel ZEB1-upregulated protein PRTG induced by Helicobacter pylori infection promotes gastric carcinogenesis through the cGMP/PKG signaling pathway.

doi: 10.1038/s41419-021-03440-1

Figure Lengend Snippet: Fig. 5 PRTG activates the downstream cGMP/PKG signaling pathway in gastric cancer cells. A Expression of top 500 genes positively associated with PRTG expression in gastric patients from TCGA database. B Gene ontology term enrichment analysis for top 8 biological process controlled by differentially expressed genes in gastric patients. C and D The expression of cGMP/PKG signaling pathway related proteins in PRTG- overexpressing AGS cells were detected by qRT-PCR (C) and western blot (D). E Supernatant cGMP levels in PRTG overexpressing AGS cells were detected by ELISA. F PRTG silencing AGS cells were infected with H. pylori for 48 h, and then the expression of cGMP/PKG signaling pathway related proteins were detected by western blot. G AGS cells were transiently co-transfected with ZEB1 overexpressing plasmid and PRTG siRNA. 48 h later, the expression of cGMP/PKG signaling pathway related proteins were detected by western blot. H–K PRTG overexpressing AGS cells were treated with sGC inhibitor (NS-2028, 10 μM) or PKG inhibitor (KT5823, 1 μM) for 24 h. cGMP levels in supernatant and phosphorylated VASP expression were detected by ELISA and western blot, respectively. Data were presented as mean ± SD from the three independent replicates. ***P < 0.0001.

Article Snippet: The following primary antibodies were used: anti-PRTG antibody (CF501394, OriGene), ZEB1(21544-1-AP, Proteintech), E-Cadherin (13-1700, Invitrogen), N-Cadherin (33-3900, Invitrogen), Vimentin (ab92547, abcam), Snail (26183-1-AP, Proteintech), sGC (ab189176, abcam), PKG1 (3248, CST), PKG2 (55138-1-AP, Proteintech), PDE5A (ab28761, abcam), VASP (13472-1-AP, Proteintech), PhosphoVASPSer239 (ab194747, abcam), GAPDH (sc-47724, Santacruz), Caspase-3 (19677-1-AP, Proteintech), p21 (10355-1-AP, Proteintech), Bid (10988-1-AP, Proteintech), BCL2 (12789-1-AP, Proteintech), PhosphoH2AXSer139 (2577, CST), BIRC3 (24304-1-AP, Proteintech), HDAC1 (sc-81598, Santa Cruz), Ubiquitin (10201-2-AP, Proteintech).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, Infection, Transfection, Plasmid Preparation

Journal: bioRxiv

Article Title: MeCP2 NID interaction with RNA: Implications for Rett Syndrome-Relevant Protein Regulation

doi: 10.1101/2025.11.19.689340

Figure Lengend Snippet: In the presence of RNA, binding to certain proteins, such as TBLR1 and Sin3a, is limited, however there are other unknown protein complexes to which MeCP2 interacts through, perhaps, scaffolding RNAs that may bind at the RBD, at some other region of the protein, or both. When RNA is degraded, some large complexes disassociate from MeCP2, as indicated from sucrose density ultracentrifugation, but MeCP2’s total protein interactome increases through interactions at the RBD, such as to TBLR1, as shown by IP data. When UV crosslinking is applied to create a covalent link between RNA bound to MeCP2, RNase treatment does not result in the increased interactome or TBLR1 binding as before, but instead obstructs TBLR1 binding, allowing other proteins, like Sin3a, to bind MeCP2 at a domain outside the RBD.

Article Snippet: Primary antibodies used were: MeCP2 (M9317, Sigma®, St. Louis, MO, USA), H4 (rabbit serum produced in-house), Sin3a (ProteinTech 14638-1-AP), TBLR1 (Bethyl, A300-408A), HDAC1 (Santa Cruz Biotechnology, sc-81598).

Techniques: RNA Binding Assay, Scaffolding, Binding Assay

Journal: Cell Death & Disease

Article Title: The novel ZEB1-upregulated protein PRTG induced by Helicobacter pylori infection promotes gastric carcinogenesis through the cGMP/PKG signaling pathway

doi: 10.1038/s41419-021-03440-1

Figure Lengend Snippet: A EdU assay after PRTG stable overexpression or transient knockdown in AGS cells. B Cell apoptosis was detected in PRTG overexpressing or silencing AGS cells after treated with chemotherapy drugs (paclitaxel, 20 nM; CDDP, 5 μM) for 48 h. C Statistical analysis of apoptosis in AGS cells ( n = 3, related to Fig. 2B). D Western blot was used to detect the effect of PRTG on the expression of apoptosis-related proteins in paclitaxel-treated AGS cells of C . E Transwell assay was used to detect the invasion and migration ability of PRTG overexpressing or silencing AGS cells. F AGS cells stably expressing empty vector (pLVX) or pLVX-PRTG were injected into the nude mice ( n = 12) and tumor volumes were monitored. G Tumor volumes of 3 mice were presented 25 days post inoculation. H Tumor volumes and weights ( n = 12) were measured 25 days post inoculation. I The expression of Ki67, E-cadherin and N-cadherin were detected by IHC. Data were presented as mean ± SD from the three independent replicates. ** P < 0.01; *** P < 0.0001. CDDP, cisplatin; pLVX, empty control pLVX lentivirus.

Article Snippet: The following primary antibodies were used: anti-PRTG antibody (CF501394, OriGene), ZEB1(21544-1-AP, Proteintech), E-Cadherin (13-1700, Invitrogen), N-Cadherin (33-3900, Invitrogen), Vimentin (ab92547, abcam), Snail (26183-1-AP, Proteintech), sGC (ab189176, abcam), PKG1 (3248, CST), PKG2 (55138-1-AP, Proteintech), PDE5A (ab28761, abcam), VASP (13472-1-AP, Proteintech), Phospho-VASP Ser239 (ab194747, abcam), GAPDH (sc-47724, Santacruz), Caspase-3 (19677-1-AP, Proteintech), p21 (10355-1-AP, Proteintech), Bid (10988-1-AP, Proteintech), BCL2 (12789-1-AP, Proteintech), Phospho-H2AX Ser139 (2577, CST), BIRC3 (24304-1-AP, Proteintech), HDAC1 (sc-81598, Santa Cruz), Ubiquitin (10201-2-AP, Proteintech).

Techniques: EdU Assay, Over Expression, Western Blot, Expressing, Transwell Assay, Migration, Stable Transfection, Plasmid Preparation, Injection