Journal: Developmental Cell

Article Title: Sestrin2 drives ER-phagy in response to protein misfolding

doi: 10.1016/j.devcel.2024.07.004

Figure Lengend Snippet:

Article Snippet: Cells were incubated in a humid chamber for 1 hour at room temperature with primary antibodies: Lamp1 (Abcam ab24170 1:200); CLIMP63 (Proteintech 16686-1-AP 1:200); HA (BioLegend 901501 1:500); Lamp1 (Santa Cruz Biotechnology sc-19992 1:500); Reep5 (Proteintech, 14643-1-AP, 1:200), CANX (Enzo Life Sciences ADI-SPA-860-D, 1:200); Flag M2 (Sigma, F1804 1:200).

Techniques: Virus, Plasmid Preparation, Retroviral, Recombinant, Cloning, Transfection, Bicinchoninic Acid Protein Assay, Mutagenesis, Control, Western Blot, Stable Transfection, Expressing, Isolation, Sequencing, Software, Microscopy, Imaging, Diagnostic Assay

Journal: Structure (London, England : 1993)

Article Title: Functional Relevance of IL-1 Receptor Inter-domain Flexibility for Cytokine Binding and Signaling

doi: 10.1016/j.str.2019.05.011

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit Anti-IL-1RAcP Antibody , Rockland , Cat# 600–401-BV3; RRID:AB_2613201.

Techniques: Recombinant, Plasmid Preparation, Software

Journal: bioRxiv

Article Title: Estradiol mediates stress-susceptibility in the male brain

doi: 10.1101/2022.01.09.475485

Figure Lengend Snippet: (A) Timeline and schematics of the subthreshold social defeat stress (SSDS) and behavioral paradigms. Male estrogen receptor alpha knockout mice demonstrated (B) social interaction deficits but not (C) anhedonia following SSDS. Male ERβ knockout mice demonstrated (D) social interaction deficits and (E) anhedonia following SSDS. Orchiectomized (ORX) mice, characterized by the absence of testosterone and consequently estradiol (E2), developed (F) social interaction deficits and (G) anhedonia following SSDS. (H-J) A single s.c. administration of E2 was effective in preventing social interaction deficits and anhedonia when administered 45 min prior to SSDS, while an administration of E2 5 min after SSDS reversed only the observed anhedonia. (K) rAAV-DIO-tdTomato injected in the nucleus accumbens (NAc) of ERβ-cre mice reveals a strong ERβ projection from the basolateral amygdala (BLA). (L-N) Whole-cell patch clamp measurement of optically-induced postsynaptic currents (oPSC) in the NAc of ERβ-cre mice that received an injection of ChR2-DIO-eYFP in the BLA. oPSCs were blocked with bath applied NBQX and APV, indicating that the ERβ-expressing BLA-NAc neurons are glutamatergic. Data shown are the mean ± S.E.M. * p <0.05; ** p <0.01; *** p <0.001. Abbreviations: artificial cerebrospinal fluid, aCSF; (2R)-amino-5-phosphonovaleric acid- APV; basolateral amygdala- BLA; dorsal,D; 17β-Estradiol, E2; estrogen receptor alpha, ERα; estrogen receptor beta, ERβ; female urine- FU; female urine sniffing test, FUST; lateral, L; medial, M; male urine, MU; nucleus accumbens, NAc; 2,3-Dioxo-6-nitro-1,2,3,4-tetrahydrobenzo[ f ]quinoxaline-7-sulfonamide, NBQX; orchiectomy, ORX; optical postsynaptic currents, oPSCs; social interaction, SI; subthreshold social defeat stress, SSDS, ventral, V.

Article Snippet: For the optogenetic (AAV5.EF1a-DIO-eYFP-WPRE-hGH or AAV5.EF1a-DIO-hChR2(H134R)-eYFP-WPRE-hGH; Penn Vector Core and Addgene) and fiber photometric (AAV5.Syn-Flex-GCaMP6S-WPRE-SV40; Addgene) experiments, mice received 400-nl bilateral injections in the basolateral amygdala (AP: −1.34, ML: 3.0, DV −5.5 from the top of the skull).

Techniques: Knock-Out, Injection, Patch Clamp, Expressing

Journal: bioRxiv

Article Title: Estradiol mediates stress-susceptibility in the male brain

doi: 10.1101/2022.01.09.475485

Figure Lengend Snippet: (A) Representative images and (B) quantification of RNAscope for Esr2 and cre in DAPI-labelled cells from the NAc and the BLA in male heterozygous ERβ mice. To confirm predominant cre expression on ERβ cells, brains from male heterozygous ERβ mice were fresh frozen and processed for RNAscope. Analysis for the expression of cre on Esr2 + cells was performed. Abbreviations: basolateral amygdala, BLA; nucleus accumbens, NAc

Article Snippet: For the optogenetic (AAV5.EF1a-DIO-eYFP-WPRE-hGH or AAV5.EF1a-DIO-hChR2(H134R)-eYFP-WPRE-hGH; Penn Vector Core and Addgene) and fiber photometric (AAV5.Syn-Flex-GCaMP6S-WPRE-SV40; Addgene) experiments, mice received 400-nl bilateral injections in the basolateral amygdala (AP: −1.34, ML: 3.0, DV −5.5 from the top of the skull).

Techniques: Expressing

Journal: bioRxiv

Article Title: Estradiol mediates stress-susceptibility in the male brain

doi: 10.1101/2022.01.09.475485

Figure Lengend Snippet: (A) Representative images, (B) quantification of RNAscope for Esr2 (transcript coding for ERβ) and retrograde tracer positive cells and (C) quantification of RNAscope for Esr2 (transcript coding for ERβ) and retrograde tracer positive cells and total BLA cells. Male wildtype mice received a conjugated cholera toxin (tracer) in the NAc. Following 2 weeks, the brains were fresh frozen and processed for Esr2 expression and colocalization with tracer + cells. Abbreviations: basolateral amygdala-,BLA; nucleus accumbens, NAc

Article Snippet: For the optogenetic (AAV5.EF1a-DIO-eYFP-WPRE-hGH or AAV5.EF1a-DIO-hChR2(H134R)-eYFP-WPRE-hGH; Penn Vector Core and Addgene) and fiber photometric (AAV5.Syn-Flex-GCaMP6S-WPRE-SV40; Addgene) experiments, mice received 400-nl bilateral injections in the basolateral amygdala (AP: −1.34, ML: 3.0, DV −5.5 from the top of the skull).

Techniques: Expressing

Journal: bioRxiv

Article Title: Estradiol mediates stress-susceptibility in the male brain

doi: 10.1101/2022.01.09.475485

Figure Lengend Snippet: (A) rAAV-DIO-tdTomato was injected in the NAc of male ERβ-cre mice and retrogradely transported to cell bodies localized in the BLA. Orchiectomized (ORX) mice received estradiol (E2; s.c.) 45 min prior to subthreshold social defeat stress (SSDS). (B) Representative immunofluorescence images from the BLA showing the co-localization of c-Fos and tdTomato in mice who received either E2 or vehicle (sesame oil) administration. (C) Increased activation of ERβ-expressing BLA to NAc cells in the anterior (A), medial (M), and posterior (P) BLA was observed in mice that received E2 prior to SSDS. (D) Schematic of the ChR2-DIO-eYFP injection in the BLA, optic fiber implantation in the NAc, and (E) representative images from the injection and fiber implantation site. (F-G) Blue light stimulation (470 nm; 20 Hz) of ERβ-expressing BLA to NAc terminals induced a real-time place preference, shown by the increased time spent in the stimulation-paired compartment. (H) Schematic of the ChR2-DIO-eYFP injection in the BLA and optic fiber implantation in the NAc. Blue light stimulation (470 nm; 4Hz) of ERβ-expressing BLA to NAc terminals for 45 min prior to stress in ORX mice prevented (I) social interaction deficits but not (J) anhedonia. Abbreviations: Anterior, A; basolateral amygdala, BLA; channelrhodopsin-2, ChR2; 17β-Estradiol, E2; estrogen receptor beta, ERβ, immunohistochemistry, IHC; middle, M; nucleus accumbens, NAc; orchiectomy, ORX; posterior, P; subthreshold social defeat stress, SSDS.

Article Snippet: For the optogenetic (AAV5.EF1a-DIO-eYFP-WPRE-hGH or AAV5.EF1a-DIO-hChR2(H134R)-eYFP-WPRE-hGH; Penn Vector Core and Addgene) and fiber photometric (AAV5.Syn-Flex-GCaMP6S-WPRE-SV40; Addgene) experiments, mice received 400-nl bilateral injections in the basolateral amygdala (AP: −1.34, ML: 3.0, DV −5.5 from the top of the skull).

Techniques: Injection, Immunofluorescence, Activation Assay, Expressing, Immunohistochemistry

Journal: bioRxiv

Article Title: Estradiol mediates stress-susceptibility in the male brain

doi: 10.1101/2022.01.09.475485

Figure Lengend Snippet: (A) Schematic of the GCaMP6s-FLEX injection in the BLA, optic fiber implantation in the NAc, and (B) representative images from the injection and implantation sites. (C) Representative traces from the fiber photometry recordings of ERβ-expressing BLA to NAc terminals in which each horizontal arrow represents an interaction bout. (D) Heatmap representation of the changes in z-score fluorescence when gonadally intact and ORX mice were interacting with a stranger or an empty cage. (E-G) Increased calcium transients were observed in ERβ-expressing BLA to NAc terminals while gonadally-intact mice were interacting with the stranger mouse. In contrast, no difference was observed in calcium activity during social interaction in ORX mice. (H) Increased calcium activity was also observed in anticipation of social interaction only in gonadally intact mice. (I) Decreased in peak width calcium activity was observed during social interaction in ORX mice (J-K) Calcium activity in gonadally-intact mice during interaction with the stranger mouse was associated with higher interaction preference compared with ORX mice. Data shown are the mean ± S.E.M. * p <0.05; ** p <0.01; *** p <0.001. Abbreviations: anticipation, Ant; basolateral amygdala, BLA; baseline, BS; estrogen receptor beta, ERβ, interaction, Int; nucleus accumbens, NAc; orchiectomy, ORX; subthreshold social defeat stress, SSDS.

Article Snippet: For the optogenetic (AAV5.EF1a-DIO-eYFP-WPRE-hGH or AAV5.EF1a-DIO-hChR2(H134R)-eYFP-WPRE-hGH; Penn Vector Core and Addgene) and fiber photometric (AAV5.Syn-Flex-GCaMP6S-WPRE-SV40; Addgene) experiments, mice received 400-nl bilateral injections in the basolateral amygdala (AP: −1.34, ML: 3.0, DV −5.5 from the top of the skull).

Techniques: Injection, Expressing, Fluorescence, Activity Assay

Journal: bioRxiv

Article Title: Estradiol mediates stress-susceptibility in the male brain

doi: 10.1101/2022.01.09.475485

Figure Lengend Snippet: Dose-response following chronic administration of estradiol (E2) reversed the orchiectomy (ORX) and SSDS-induced (C) social interaction deficits and (D) anhedonia. (C) Conversion of a brain selective E2 prodrug DHED to E2. (D) Intact C57Bl/6J male mice that received d3-DHED revealed that DHED derived E2 is found enriched in the brain but only in limited quantities in serum. Chronic administration of DHED with minipumps prevented the SSDS/orchiectomy- induced (E) social interaction deficits and (F) anhedonia. (G) Schematic representation and summary of the suggested mechanism for stress susceptibility in male mice. Data shown are the mean ± S.E.M. * p <0.05; ** p <0.01; *** p <0.001. basolateral amygdala, BLA ; (17β)-10,17-Dihydroxy-estra-1,4-dien-3-one, DHED; 17β-Estradiol, E2; estrogen receptor beta, ERβ; orchiectomy, ORX; nucleus accumbens, NAc.

Article Snippet: For the optogenetic (AAV5.EF1a-DIO-eYFP-WPRE-hGH or AAV5.EF1a-DIO-hChR2(H134R)-eYFP-WPRE-hGH; Penn Vector Core and Addgene) and fiber photometric (AAV5.Syn-Flex-GCaMP6S-WPRE-SV40; Addgene) experiments, mice received 400-nl bilateral injections in the basolateral amygdala (AP: −1.34, ML: 3.0, DV −5.5 from the top of the skull).

Techniques: Derivative Assay

Journal: Molecular Medicine Reports

Article Title: Effects of cyclooxygenase-2 gene silencing on the biological behavior of SKOV3 ovarian cancer cells

doi: 10.3892/mmr.2014.2732

Figure Lengend Snippet: Sequencing chromatogram of the pGPU6-small hairpin RNA-cyclooxygenase-2 recombinant plasmid.

Article Snippet: The positive recombinant plasmids were identified by Shanghai Yingjun Biotechnology Co., Ltd. (Shanghai, China).

Techniques: Sequencing, Recombinant, Plasmid Preparation

Journal: Molecular Medicine Reports

Article Title: Effects of cyclooxygenase-2 gene silencing on the biological behavior of SKOV3 ovarian cancer cells

doi: 10.3892/mmr.2014.2732

Figure Lengend Snippet: Assessment of transfection efficiency in SKOV3 cells transfected with recombinant plasmid (magnification, ×20). (A1 and 2) Fluorescent and transmitted light images of SKOV3 cells transfected with pGPU6-cyclooxygenase-2-shRNA recombinant plasmid. (B1 and 2) Fluorescent and transmitted light images of SKOV3 cells transfected with pGPU6-Control-shRNA recombinant plasmid. sh, small hairpin.

Article Snippet: The positive recombinant plasmids were identified by Shanghai Yingjun Biotechnology Co., Ltd. (Shanghai, China).

Techniques: Transfection, Recombinant, Plasmid Preparation, shRNA, Control

Journal: iScience

Article Title: Chidamide suppresses adipogenic differentiation of bone marrow derived mesenchymal stem cells via increasing REEP2 expression

doi: 10.1016/j.isci.2023.106221

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-REEP2 , Proteintech , Cat#15684-1-AP; RRID: AB_2238326.

Techniques: Recombinant, SYBR Green Assay, Protease Inhibitor, Stripping Membranes, cDNA Synthesis, Chromatin Immunoprecipitation, Bicinchoninic Acid Protein Assay, Plasmid Preparation, Purification, Expressing, Software

Journal: ACS Omega

Article Title: SARS-CoV-2 Protein Nsp9 Is Involved in Viral Evasion through Interactions with Innate Immune Pathways

doi: 10.1021/acsomega.4c02631

Figure Lengend Snippet: SARS-CoV-2 Nsp9 Western blot analysis. A) SARS-CoV-2 Orf6, Orf3a, and Nsp9 expression. A549 cells were reverse transfected with 2 μg Orf6, Orf3a, or Nsp9 expression vectors and then lysed for protein detection by Western blotting. B) Western blotting of p-NF-kB, NF-kB, and TBK1 protein expression in 24 h poly I:C-treated A549 cells transfected with Nsp9 or empty vector control plasmids. B-tubulin is used as a control for protein expression. C) Densitometry analysis showing relative TBK1 abundance in 24 h poly I:C (PIC)-treated A549 cells transfected with Nsp9 or empty vector control plasmids compared to nontransfected and nontreated A549 cells ( n = 3). Error bars represent standard error of the mean, * = p < 0.05.

Article Snippet: SARS-CoV-2 nonstructural and accessory protein expression vectors were purchased from Addgene (MA, USA): pTwist-CMV-Nsp9–3xFlag (pGBW- m4133209), pTwist-CMV-ORF6–3xFlag (pGBW-m4252831), and pTwist-CMV-ORF3a- 3xFlag (pGBW-m4133902).

Techniques: Western Blot, Expressing, Transfection, Plasmid Preparation, Control

Journal: Cell Communication and Signaling : CCS

Article Title: CLCA1 suppresses colorectal cancer aggressiveness via inhibition of the Wnt/beta-catenin signaling pathway

doi: 10.1186/s12964-017-0192-z

Figure Lengend Snippet: Decreased expression level of CLCA1 in CRC tissues. a Representative images of IHC staining of CLCA1 in paraffin-embedded CRC tissues and matched adjacent normal mucosa. b IHC staining scores in the CRC group and adjacent normal tissues ( P < 0.0001, paired t -test). c Representative bands from western blotting of matched fresh-frozen CRC and adjacent normal mucosa. d Relative CLCA1 protein intensity in the CRC group and adjacent normal tissues ( P < 0.0001, paired t -test). Abbreviations: IHC, immunohistochemistry; GAPDH, glyceraldehyde-3-phosphate dehydrogenase

Article Snippet: CLCA1 CRISPR/Cas9 (the Clustered Regularly Interspaced Short Palindromic Repeats and CRISPR-associated protein system) activation plasmid (sc-402,998-ACT) and CLCA1 CRISPR/Cas9 knockout plasmid (double nickase plasmid, sc-402,998-NIC) were bought from Santa Cruz Biotechnology and transfected into SW620 cells according to the manufacturer’s protocols.

Techniques: Expressing, Immunohistochemistry, Western Blot

Journal: Cell Communication and Signaling : CCS

Article Title: CLCA1 suppresses colorectal cancer aggressiveness via inhibition of the Wnt/beta-catenin signaling pathway

doi: 10.1186/s12964-017-0192-z

Figure Lengend Snippet: CLCA1 serum concentration and CLCA1 mRNA expression level are inversely correlated with CRC metastasis. a ELISA results showed that the CLCA1 serum expression level was significantly lower in CRC patients than in healthy controls (1.48 ± 1.06 ng/mL vs 1.06 ± 0.73 ng/mL, P = 0.0018, t -test). b Compared to that in patients with early stage disease (TNM stage I and II), the CLCA1 serum expression level was markedly reduced in patients with local or distant metastasis (TNM stage III and IV) (1.24 ± 0.86 ng/mL vs 0.88 ± 0.53 ng/mL, P = 0.013, t -test). c CLCA1 mRNA level in CRC samples from TCGA database. Abbreviations : ELISA, enzyme-linked immunosorbent assay; TCGA, the Cancer Genome Atlas

Article Snippet: CLCA1 CRISPR/Cas9 (the Clustered Regularly Interspaced Short Palindromic Repeats and CRISPR-associated protein system) activation plasmid (sc-402,998-ACT) and CLCA1 CRISPR/Cas9 knockout plasmid (double nickase plasmid, sc-402,998-NIC) were bought from Santa Cruz Biotechnology and transfected into SW620 cells according to the manufacturer’s protocols.

Techniques: Concentration Assay, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Cell Communication and Signaling : CCS

Article Title: CLCA1 suppresses colorectal cancer aggressiveness via inhibition of the Wnt/beta-catenin signaling pathway

doi: 10.1186/s12964-017-0192-z

Figure Lengend Snippet: Increased expression level of CLCA1 suppresses CRC cell growth in vitro. a Western blotting analysis of CLCA1 in the SW620 cell line transfected with CLCA1 CRISPR/Cas9-activation and -knockout plasmids. b qRT-PCR analysis of CLCA1 in the SW620 cell line transfected with CLCA1 CRISPR/Cas9-activation plasmids. c qRT-PCR analysis of CLCA1 in SW620 cell line transfected with CLCA1 CRISPR/Cas9-knockout plasmids. d Growth curves of the CLCA1-ACT and CLCA1-ACT-NC group. e Growth curves of the CLCA1-KO and CLCA1-KO-NC group. Abbreviations: qRT-PCR, quantitative reverse transcription polymerase chain reaction; CRISPR/Cas9, the Clustered Regularly Interspaced Short Palindromic Repeats and CRISPR-associated protein system; CLCA1-ACT, CLCA1-upregulated cells; CLCA1-ACT-NC, negative control of CLCA1-upregulated cells; CLCA1-KO, CLCA1-knockout cells; CLCA1-KO-NC, negative control of CLCA1-knockout cells. * P < 0.05, t -test; *** P < 0.001, t -test

Article Snippet: CLCA1 CRISPR/Cas9 (the Clustered Regularly Interspaced Short Palindromic Repeats and CRISPR-associated protein system) activation plasmid (sc-402,998-ACT) and CLCA1 CRISPR/Cas9 knockout plasmid (double nickase plasmid, sc-402,998-NIC) were bought from Santa Cruz Biotechnology and transfected into SW620 cells according to the manufacturer’s protocols.

Techniques: Expressing, In Vitro, Western Blot, Transfection, CRISPR, Activation Assay, Knock-Out, Quantitative RT-PCR, Reverse Transcription, Polymerase Chain Reaction, Negative Control

Journal: Cell Communication and Signaling : CCS

Article Title: CLCA1 suppresses colorectal cancer aggressiveness via inhibition of the Wnt/beta-catenin signaling pathway

doi: 10.1186/s12964-017-0192-z

Figure Lengend Snippet: Increased expression level of CLCA1 suppresses CRC cell colony formation ability in vitro. a Images of the colony formation assay with CLCA1-ACT and CLCA1-ACT-NC cells. b The mean number of colonies in CLCA1-ACT and CLCA1-ACT-NC cells ( P < 0.0001 t -test). c Images of the colony formation assay with CLCA1-KO and CLCA1-KO-NC cells. d Mean number of colonies in the CLCA1-KO and CLCA1-KO-NC cells ( P < 0.0001, t -test). Abbreviations: CLCA1-ACT, CLCA1-upregulated cells; CLCA1-ACT-NC, negative control of CLCA1-upregulated cells; CLCA1-KO, CLCA1-knockout cells; CLCA1-KO-NC, negative control of CLCA1-knockout cells

Article Snippet: CLCA1 CRISPR/Cas9 (the Clustered Regularly Interspaced Short Palindromic Repeats and CRISPR-associated protein system) activation plasmid (sc-402,998-ACT) and CLCA1 CRISPR/Cas9 knockout plasmid (double nickase plasmid, sc-402,998-NIC) were bought from Santa Cruz Biotechnology and transfected into SW620 cells according to the manufacturer’s protocols.

Techniques: Expressing, In Vitro, Colony Assay, Negative Control, Knock-Out

Journal: Cell Communication and Signaling : CCS

Article Title: CLCA1 suppresses colorectal cancer aggressiveness via inhibition of the Wnt/beta-catenin signaling pathway

doi: 10.1186/s12964-017-0192-z

Figure Lengend Snippet: Increased expression level of CLCA1 reduces CRC cell migration and invasion in vitro. a Cell migration and invasion assays in CLCA1-ACT and CLCA1-ACT-NC cells (left panel, representative pictures of transwell chambers; right panel, average counts of five random microscopic fields at a magnification of 200×). b Cell migration and invasion assays in CLCA1-KO and CLCA1-KO-NC cells (left panel, representative pictures of transwell chambers; right panel, average counts of five random microscopic fields at a magnification of 200×)

Article Snippet: CLCA1 CRISPR/Cas9 (the Clustered Regularly Interspaced Short Palindromic Repeats and CRISPR-associated protein system) activation plasmid (sc-402,998-ACT) and CLCA1 CRISPR/Cas9 knockout plasmid (double nickase plasmid, sc-402,998-NIC) were bought from Santa Cruz Biotechnology and transfected into SW620 cells according to the manufacturer’s protocols.

Techniques: Expressing, Migration, In Vitro

Journal: Cell Communication and Signaling : CCS

Article Title: CLCA1 suppresses colorectal cancer aggressiveness via inhibition of the Wnt/beta-catenin signaling pathway

doi: 10.1186/s12964-017-0192-z

Figure Lengend Snippet: Increased expression level of CLCA1 inhibits CRC proliferation and metastasis in vivo. a Subcutaneous xenograft mouse models (arrows show subcutaneous tumors). b Dissected subcutaneous xenograft tumors. c Representative H & E staining images and CLCA1 IHC staining images of xenograft tumors. d Subcutaneous xenograft tumor growth curves of the CLCA1-ACT and CLCA1-ACT-NC group. e Average tumor weight of the CLCA1-ACT and CLCA1-ACT-NC group. Abbreviations: H & E, Hematoxylin and eosin; IHC, immunohistochemistry

Article Snippet: CLCA1 CRISPR/Cas9 (the Clustered Regularly Interspaced Short Palindromic Repeats and CRISPR-associated protein system) activation plasmid (sc-402,998-ACT) and CLCA1 CRISPR/Cas9 knockout plasmid (double nickase plasmid, sc-402,998-NIC) were bought from Santa Cruz Biotechnology and transfected into SW620 cells according to the manufacturer’s protocols.

Techniques: Expressing, In Vivo, Staining, Immunohistochemistry

Journal: Cell Communication and Signaling : CCS

Article Title: CLCA1 suppresses colorectal cancer aggressiveness via inhibition of the Wnt/beta-catenin signaling pathway

doi: 10.1186/s12964-017-0192-z

Figure Lengend Snippet: Increased expression level of CLCA1 inhibits CRC metastasis in vivo. a Representative liver with metastatic tumors (arrows) from the CLCA1-ACT-NC group. b Representative liver without visible metastatic tumors from the CLCA1-ACT group. c H & E staining image of liver metastasis in the CLCA1-ACT group. d Average number of visible liver metastatic nodules

Article Snippet: CLCA1 CRISPR/Cas9 (the Clustered Regularly Interspaced Short Palindromic Repeats and CRISPR-associated protein system) activation plasmid (sc-402,998-ACT) and CLCA1 CRISPR/Cas9 knockout plasmid (double nickase plasmid, sc-402,998-NIC) were bought from Santa Cruz Biotechnology and transfected into SW620 cells according to the manufacturer’s protocols.

Techniques: Expressing, In Vivo, Staining

Journal: Cell Communication and Signaling : CCS

Article Title: CLCA1 suppresses colorectal cancer aggressiveness via inhibition of the Wnt/beta-catenin signaling pathway

doi: 10.1186/s12964-017-0192-z

Figure Lengend Snippet: CLCA1 expression level upregulation inhibits EMT and Wnt/beta-catenin signaling. a Western blotting analysis showed that the increased expression level of CLCA1 increased the E-cadherin expression level but repressed N-cadherin, vimentin, slug, and snail expression levels. CLCA1 expression level downregulation caused the reverse effects. b Beta-catenin nuclear translocation was decreased after CLCA1 expression level was upregulated and increased after CLCA1 was knocked down. c The expression levels of beta-catenin and mesenchymal markers were downregulated in CLCA1-KO cells after treatment with XAV939. Abbreviations : EMT, epithelial-mesenchymal transition

Article Snippet: CLCA1 CRISPR/Cas9 (the Clustered Regularly Interspaced Short Palindromic Repeats and CRISPR-associated protein system) activation plasmid (sc-402,998-ACT) and CLCA1 CRISPR/Cas9 knockout plasmid (double nickase plasmid, sc-402,998-NIC) were bought from Santa Cruz Biotechnology and transfected into SW620 cells according to the manufacturer’s protocols.

Techniques: Expressing, Western Blot, Translocation Assay

Journal: Cell Communication and Signaling : CCS

Article Title: CLCA1 suppresses colorectal cancer aggressiveness via inhibition of the Wnt/beta-catenin signaling pathway

doi: 10.1186/s12964-017-0192-z

Figure Lengend Snippet: Promotion of CRC cell aggressiveness was abrogated when CLCA1-KO cells were co-treated with XAV939. a The promotion of CRC cell growth was abrogated when CLCA1-KO cells were co-treated with XAV939. b The enhancement CRC cell migration and invasion was abrogated when CLCA1-KO cells were co-treated with XAV939 (left panel, representative pictures of transwell chambers; right panel, average counts of five random microscopic fields at a magnification of 200×). * P < 0.05, t -test; ** P < 0.01, t -test

Article Snippet: CLCA1 CRISPR/Cas9 (the Clustered Regularly Interspaced Short Palindromic Repeats and CRISPR-associated protein system) activation plasmid (sc-402,998-ACT) and CLCA1 CRISPR/Cas9 knockout plasmid (double nickase plasmid, sc-402,998-NIC) were bought from Santa Cruz Biotechnology and transfected into SW620 cells according to the manufacturer’s protocols.

Techniques: Migration