Journal: Frontiers in immunology

Article Title: Gelsolin Attenuates Neonatal Hyperoxia-Induced Inflammatory Responses to Rhinovirus Infection and Preserves Alveolarization.

doi: 10.3389/fimmu.2022.792716

Figure Lengend Snippet: FIGURE 1 | In neonatal mice, gelsolin blocks hyperoxia-induced pro-inflammatory responses to RV infection. Two-day old wild type mice were exposed to hyperoxia or normoxia for 10 days and treated with recombinant human plasma gelsolin (GSN, 0.5 mg/kg) or equal volume BSA-PBS, administered intranasally under anesthesia daily. On day of life 14 the mice were inoculated with RV. Whole lung mRNA expression of Il12p40, Tnfa and Ifng (A) and protein expression of Il12p70, TNF-a and IFN-g (B) were measured 2 days later. (N = 4-6, mean ± SEM, *p < 0.05, **p < 0.01, NS, nonsignificant, ANOVA). One of two independent experiments is shown.

Article Snippet: In selected experiments, neonatal mice were treated with 10ul of PBS (Sigma D6662) or recombinant human plasma gelsolin (Cytoskeleton Inc., HPG6) 0.5mg/kg intranasally daily during exposure to hyperoxia.

Techniques: Infection, Recombinant, Clinical Proteomics, Expressing

Journal: Frontiers in immunology

Article Title: Gelsolin Attenuates Neonatal Hyperoxia-Induced Inflammatory Responses to Rhinovirus Infection and Preserves Alveolarization.

doi: 10.3389/fimmu.2022.792716

Figure Lengend Snippet: FIGURE 2 | Gelsolin inhibits hyperoxia-induced CD103+ DC expansion and inflammation. Two-day old wild type mice were exposed to hyperoxia or normoxia for 4 or 10 days and treated with recombinant human plasma gelsolin (GSN, 0.5 mg/kg) or equal volume BSA-PBS, administered intranasally under anesthesia daily. Lungs were enzymatically digested, and a single cell suspension was incubated and stained with specific cell-surface antibodies. Lung CD103+ DCs were distinguished from other lung cells based on expression of CD45, F4/80, CD11c, CD103 and CD11b. (A) Gating strategy to identify lung DCs. (B) Conventional lung DC populations are distinguished based on the expression of CD103 and CD11b. Following hyperoxia, two distinct populations of CD103+ DCs, CD103lo and CD103hi DCs are observed. GSN treatment during hyperoxia decreases CD103lo, but not CD103hi DCs. (C) Quantification of the CD103lo and CD103hi DCs. (D) Both CD103lo and CD103hi DC populations are absent in the lungs of 4-day old Batf3 null mice compared to age-matched wild type (WT) mice. *p < 0.05, **p < 0.01, ***p < 0.001, NS, nonsignificant (ANOVA). These results are representative of three independent experiments. (E) RNA was extracted from the whole lung tissue on day of life 16 after 10 days exposure to normoxia or hyperoxia with and without daily GSN treatment. GSN attenuated hyperoxia-induced mRNA expression of Il12p40, Myd88, Cd207, Cd103 and Clec9a. *p < 0.05, **p < 0.01 (ANOVA). N = 4 per groups. One of three independent experiments is shown.

Article Snippet: In selected experiments, neonatal mice were treated with 10ul of PBS (Sigma D6662) or recombinant human plasma gelsolin (Cytoskeleton Inc., HPG6) 0.5mg/kg intranasally daily during exposure to hyperoxia.

Techniques: Recombinant, Clinical Proteomics, Suspension, Incubation, Staining, Expressing

Journal: Frontiers in immunology

Article Title: Gelsolin Attenuates Neonatal Hyperoxia-Induced Inflammatory Responses to Rhinovirus Infection and Preserves Alveolarization.

doi: 10.3389/fimmu.2022.792716

Figure Lengend Snippet: FIGURE 3 | Gelsolin attenuates hyperoxic BALF-induced inflammatory response to RV. Two-day old mice were exposed to hyperoxia or normoxia for 14 days. BALF was collected on day of life 16. F-actin and gelsolin (GSN) protein levels were measured in BALF supernatant using ELISA and the ratio of F-actin to GSN was calculated and compared between samples from hyperoxia- and normoxia-exposed mice, unpaired t-test. *p < 0.05 (A). (B) 14-day old mice were exposed to hyperoxia for 4 days and BALF was collected (H-BALF). Cell-free H-BALF was incubated with GSN (5mg) in vitro for 10min and F-actin levels were analyzed by ELISA (paired t-test p < 0.001). (C) 14-day old mice were inoculated with H-BALF, RV, GSN, or appropriate controls and whole lung mRNA expression of Il12p40, Ifng and Tnfa were analyzed 2 days later. *p < 0.05, ***p < 0.001 (ANOVA). N = 3-6 per group. One of two independent experiments is shown. ns, not significant

Article Snippet: In selected experiments, neonatal mice were treated with 10ul of PBS (Sigma D6662) or recombinant human plasma gelsolin (Cytoskeleton Inc., HPG6) 0.5mg/kg intranasally daily during exposure to hyperoxia.

Techniques: Enzyme-linked Immunosorbent Assay, Incubation, In Vitro, Expressing

Journal: Frontiers in immunology

Article Title: Gelsolin Attenuates Neonatal Hyperoxia-Induced Inflammatory Responses to Rhinovirus Infection and Preserves Alveolarization.

doi: 10.3389/fimmu.2022.792716

Figure Lengend Snippet: FIGURE 5 | Gelsolin prevents neonatal hyperoxia-induced hypoalveolarization. Two-day old wild type mice were exposed to hyperoxia or normoxia for 10 days and treated with recombinant human plasma gelsolin (0.5 mg/kg) or equal volume BSA-PBS, administered intranasally under anesthesia daily. Lung histology was assessed on day of life 16. (A) Representative lung sections were stained with hematoxylin and eosin. Hyperoxia induced enlargement of the alveolar spaces (hypoalveolarization) in BSA-PBS-treated mice. In contrast, gelsolin treatment preserved the alveolar architecture of hyperoxia-exposed mice. (B) Alveolar chord length was significantly increased in hyperoxia-exposed, BSA-PBS-treated mice, consistent with hypoalveolarization. In contrast, alveolar chord length of hyperoxia- exposed, gelsolin-treated mice was similar to that of normoxia-exposed mice, indicating protective effect of gelsolin on alveolarization during neonatal hyperoxia. ****p < 0.0001 (ANOVA). N = 5-12 per group.

Article Snippet: In selected experiments, neonatal mice were treated with 10ul of PBS (Sigma D6662) or recombinant human plasma gelsolin (Cytoskeleton Inc., HPG6) 0.5mg/kg intranasally daily during exposure to hyperoxia.

Techniques: Recombinant, Clinical Proteomics, Staining

Journal: Frontiers in immunology

Article Title: Gelsolin Attenuates Neonatal Hyperoxia-Induced Inflammatory Responses to Rhinovirus Infection and Preserves Alveolarization.

doi: 10.3389/fimmu.2022.792716

Figure Lengend Snippet: FIGURE 6 | F-actin/Gelsolin ratio in tracheal aspirates of mechanically ventilated human preterm infants with respiratory distress and FiO2 on day of sample collection. Tracheal aspirates were collected in the first week of life. F-actin and gelsolin levels were measured by ELISA in the supernatant. The association between the ratio of F-actin to gelsolin and the FiO2 on the day of sample collection was determined by Pearson’s correlation analysis.

Article Snippet: In selected experiments, neonatal mice were treated with 10ul of PBS (Sigma D6662) or recombinant human plasma gelsolin (Cytoskeleton Inc., HPG6) 0.5mg/kg intranasally daily during exposure to hyperoxia.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Frontiers in immunology

Article Title: Gelsolin Attenuates Neonatal Hyperoxia-Induced Inflammatory Responses to Rhinovirus Infection and Preserves Alveolarization.

doi: 10.3389/fimmu.2022.792716

Figure Lengend Snippet: FIGURE 7 | In tracheal aspirates of human preterm infants with respiratory distress, gelsolin concentrations decrease during the first two weeks of mechanical ventilation. For preterm infants who remain endotracheally intubated receiving mechanical ventilation tracheal aspirates were obtained during week 1 and week 2 of mechanical ventilation and gelsolin levels in the supernatant were quantified. Statistical significance was pointed by paired t-test.

Article Snippet: In selected experiments, neonatal mice were treated with 10ul of PBS (Sigma D6662) or recombinant human plasma gelsolin (Cytoskeleton Inc., HPG6) 0.5mg/kg intranasally daily during exposure to hyperoxia.

Techniques:

Journal: Translational Oncology

Article Title: Downregulation of PIP4K2C inhibits the breast cancer cell proliferation, migration and invasion

doi: 10.1016/j.tranon.2025.102420

Figure Lengend Snippet: The mRNA expression of PIP4K2C in pan-cancer. (A) The mRNA expression of PIP4K2C in 33 tumors in TCGA GTEx samples (ns, p > 0.05; * p < 0.05; ** p < 0.01; *** p < 0.001). (B) PIP4K2C expression in the breast cancer tissues and unpaired normal samples. (C) The expression level of PIP4K2C in the breast cancer tissues and the paired normal samples. ACC, adrenocortical carcinoma; BLCA, bladder urothelial carcinoma; BRCA, breast invasive carcinoma; CESC, cervical and endocervical cancers; CHOL, cholangiocarcinoma; COAD, colon adenocarcinoma; DLBC, lymphoid neoplasm diffuse large B-cell lymphoma; ESCA, esophageal carcinoma; GBM, glioblastoma multiforme; HNSC, head and neck squamous cell carcinoma; KICH, kidney chromophobe; KIRC, kidney renal clear cell carcinoma; KIRP, kidney renal papillary cell carcinoma; LAML, acute myeloid leukemia; LGG, brain lower grade glioma; LIHC, liver hepatocellular carcinoma; LUAD, lung adenocarcinoma; LUSC, lung squamous cell carcinoma; MESO, mesothelioma; OV, ovarian serous cystadenocarcinoma; PAAD, pancreatic adenocarcinoma; PCPG, pheochromocytoma and paraganglioma; PRAD, prostate adenocarcinoma; READ, rectum adenocarcinoma; SARC, sarcoma; SKCM, skin cutaneous melanoma; STAD, stomach adenocarcinoma; STES, stomach and esophageal carcinoma; TGCT, testicular germ cell tumors; THCA, thyroid carcinoma; THYM, thymoma; UCEC, uterine corpus endometrial carcinoma; UCS, uterine carcinosarcoma; UVM, uveal melanoma.

Article Snippet: In this assay, primary antibody PIP4K2C (1:100, CUSABIO, CSB-PA819455LA01HU) and secondary antibodies Goat anti-rabbit IgG H&L (Alexa Fluor® 488) (1:400, abcam, ab150077) were used.

Techniques: Expressing

Journal: Translational Oncology

Article Title: Downregulation of PIP4K2C inhibits the breast cancer cell proliferation, migration and invasion

doi: 10.1016/j.tranon.2025.102420

Figure Lengend Snippet: mRNA expression and protein levels of PIP4K2C in breast cancer cell lines and tissues. (A) The expression level of PIP4K2C in the normal mammary gland cell line MCF-10A and breast cancer cell lines (MDA-MB-231, MDA-MB-468, MCF7, ZR751 and BT20) was determined using qPCR. * P < 0.05, *** P < 0.01 vs MCF-10A. (B) The protein levels of PIP4K2C in cell lines were measured by western blot. (C) The expression level of PIP4K2C in the breast cancer tissues and the paired normal samples. (D) The protein levels of PIP4K2C in the breast cancer tissues and the paired normal samples. (E) The immunofluorescence staining of the breast cancer tissues and the paired normal samples. (F) The IHC images of PIP4K2C in normal and tumor tissues.

Article Snippet: In this assay, primary antibody PIP4K2C (1:100, CUSABIO, CSB-PA819455LA01HU) and secondary antibodies Goat anti-rabbit IgG H&L (Alexa Fluor® 488) (1:400, abcam, ab150077) were used.

Techniques: Expressing, Western Blot, Immunofluorescence, Staining

Journal: Translational Oncology

Article Title: Downregulation of PIP4K2C inhibits the breast cancer cell proliferation, migration and invasion

doi: 10.1016/j.tranon.2025.102420

Figure Lengend Snippet: PIP4K2C was knocked down by siRNA. (A-B) The transfection efficiency of MDA-MB-468 was detected at mRNA expression and protein levels, respectively. (C-D) The transfection efficiency of MCF7 was detected at mRNA expression and protein levels (48 h), respectively.

Article Snippet: In this assay, primary antibody PIP4K2C (1:100, CUSABIO, CSB-PA819455LA01HU) and secondary antibodies Goat anti-rabbit IgG H&L (Alexa Fluor® 488) (1:400, abcam, ab150077) were used.

Techniques: Transfection, Expressing

Journal: Translational Oncology

Article Title: Downregulation of PIP4K2C inhibits the breast cancer cell proliferation, migration and invasion

doi: 10.1016/j.tranon.2025.102420

Figure Lengend Snippet: PIP4K2C was overexpressed in MCF 10A by transfection. (A) The transfection efficiency was detected at mRNA expression. (B) overexpression of PIP4K2C resulted in increased proliferation.

Article Snippet: In this assay, primary antibody PIP4K2C (1:100, CUSABIO, CSB-PA819455LA01HU) and secondary antibodies Goat anti-rabbit IgG H&L (Alexa Fluor® 488) (1:400, abcam, ab150077) were used.

Techniques: Transfection, Expressing, Over Expression

Journal: Translational Oncology

Article Title: Downregulation of PIP4K2C inhibits the breast cancer cell proliferation, migration and invasion

doi: 10.1016/j.tranon.2025.102420

Figure Lengend Snippet: Inhibition of PIP4K2C suppressed the proliferation, migration and invasion of MDA-MB-468 and MCF7 cells. (A) Knockdown of PIP4K2C by siRNA resulted in reduced proliferation. (B) The reduced cell migration rate was evaluated by wound healing assay. The percentage of wound closure at 24 and 48 h was calculated using ImageJ based on the change in scratch area from time 0 h. (C-D) The cell migration and invasion ability were detected by transwell assay.

Article Snippet: In this assay, primary antibody PIP4K2C (1:100, CUSABIO, CSB-PA819455LA01HU) and secondary antibodies Goat anti-rabbit IgG H&L (Alexa Fluor® 488) (1:400, abcam, ab150077) were used.

Techniques: Inhibition, Migration, Knockdown, Wound Healing Assay, Transwell Assay

Journal: Translational Oncology

Article Title: Downregulation of PIP4K2C inhibits the breast cancer cell proliferation, migration and invasion

doi: 10.1016/j.tranon.2025.102420

Figure Lengend Snippet: Down-regulation of PIP4K2C enhanced the protein levels of LC3II/LC3I.

Article Snippet: In this assay, primary antibody PIP4K2C (1:100, CUSABIO, CSB-PA819455LA01HU) and secondary antibodies Goat anti-rabbit IgG H&L (Alexa Fluor® 488) (1:400, abcam, ab150077) were used.

Techniques:

Journal: eLife

Article Title: miR-252 targeting temperature receptor CcTRPM to mediate the transition from summer-form to winter-form of Cacopsylla chinensis

doi: 10.7554/eLife.88744

Figure Lengend Snippet: ( A ) The predicted protein tertiary structure of CcTRPM . The conserved ankyrin repeat (ANK) domain was indicated in the N-terminal. The conserved six transmembrane domain of ion channels structure were shown as TM1-TM6. ( B ) The mRNA expression of CcTRPM in response to different temperatures of 25°C and 10°C by qRT-PCR. ( C ) Fluorescence detection of Fluo-4 AM after heterologous expression of CcTRPM in mammalian HEK293T cells in response to menthol treatment. The recombinant plasmid was generated by inserting the full ORF sequence of CcTRPM into pcDNA3.1(+)-mCherry plasmid. DMSO treatment and ionomycin treatment were used as negative control and positive control, separately. ( D ) Representative images of Ca 2+ imaging after heterologous expression of CcTRPM in mammalian HEK293T cells in response to different temperature treatment. “CcTRPM +10 °C” means the recombinant plasmid of CcTRPM with pcDNA3.1(+)-mCherry was treated with 10 °C. “CcTRPM +25 °C” denotes the pcDNA3.1(+)-mCherry plasmid with CcTRPM was treated with 25 °C. mCherry and Fluo-4 signal are shown in red and green, respectively. Scale bar is 20 μm. ( E ) The developmental expression pattern of CcTRPM for SF at mRNA level using qRT-PCR. 1st, 2nd, 3rd, 4th, and 5th are the nymphs at the first, second, third, fourth and fifth instar, respectively. A-1d, A-5d, and A-10d are the adults at 1 day, 5 days, and 10 days, separately. ( F ) Representative confocal images of CcTRPM in different developmental stages of SF using FISH. Scale bar is 0.5 mm. BF: the bright field. DAPI: the cell nuclei were stained with DAPI and visualized in blue. CcTRPM-Cy3: CcTRPM signal was labeled with Cy3 and visualized in red. Merge: merged imaging of BF, DAPI, and CcTRPM-Cy3 signals. ( G–H ) Comparison of the nymph cuticle ultrastructure, cuticle thickness, and cuticle chitin content of SF 1st instar treated with dsCcTRPM and dsEGFP at 15 days. ( I ) The transition percent of SF 1st instar nymphs treated with dsEGFP, DMSO, dsCcTRPM, CcTRPM antagonist and dsCcTRPM +menthol at 15 days under 10 °C. For RNAi experiments, summer-form 1st instar nymphs were fed with dsEGFP (500 ng/μL) or dsCcTRPM (500 ng/μL). To mimic RNAi effect, summer-form 1st instar nymphs were fed with 0.1% DMSO or CcTRPM antagonist (20 ng/μL). For the rescue experiment, summer-form 1st instar nymphs were fed with the mixture of dsCcTRPM (500 ng/μL) and menthol (1 mg/mL). Then, counted the number of summer-form and winter-form individuals and calculated the transition percent. ( J ) The phenotypes of SF 1st instar nymphs treated with dsEGFP, dsCcTRPM, CcTRPM antagonist and dsCcTRPM +menthol at 15 days under 10 °C. The data in 2B and 2E are shown as the mean ± SD with three independent biological replications of at least 30 nymphs for each biological replication. Scale bar is 0.5 mm. Data in 2 G and 2 H are presented as mean ± SD with three biological replications of three technical replicates for each biological replication. Data in 2I are presented as mean ± SD with nine biological replications. Statistically significant differences were determined with the pair-wise Student’s t -test, and significance levels were denoted by *** (p<0.001). Different letters above the bars indicated statistically significant differences (p<0.05), as determined by ANOVA followed with a Turkey’s HSD multiple comparison test in SPSS 20.0 software.

Article Snippet: We therefore attempted to identify chemical compounds that either activate or inhibit CcTRPM . qRT-PCR results revealed that menthol treatment with all three concentrations significantly increased the mRNA expression of CcTRPM and antagonist (HY-112430, MedChemExpress, Shanghai, China) treatment with all three concentrations markedly reduced CcTRPM transcription ( ).

Techniques: Expressing, Quantitative RT-PCR, Fluorescence, Recombinant, Plasmid Preparation, Generated, Sequencing, Negative Control, Positive Control, Imaging, Staining, Labeling, Comparison, Software

Journal: eLife

Article Title: miR-252 targeting temperature receptor CcTRPM to mediate the transition from summer-form to winter-form of Cacopsylla chinensis

doi: 10.7554/eLife.88744

Figure Lengend Snippet: ( A–B ) The chemical structure of menthol and TRPM antagonist. ( C–E ) Melting curve for qRT-PCR primers of CcTRPM , Ccβ-actin and CcEF-1 . ( F ) The mRNA expression of CcTRPM in response to different temperatures of 25°C and 10°C at 1 day and 2 days by qRT-PCR. ( G ) Effect of menthol treatment at different concentrations on the mRNA expression of CcTRPM under 10 °C condition. ( H ) Effect of TRPM antagonist treatment at different concentrations on the mRNA expression of CcTRPM under 10 °C condition. ( I ) Representative images of Ca 2+ imaging after heterologous expression of CcTRPM in mammalian HEK293T cells in response to menthol treatment. ‘CcTRPM +menthol’ means the recombinant plasmid of CcTRPM with pcDNA3.1(+)-mCherry was treated with menthol. “CcTRPM +DMSO” denotes the pcDNA3.1(+)-mCherry plasmid with CcTRPM was treated with DMSO. mCherry and Fluo-4 signal are shown in red and green, respectively. Scale bar is 20 μm. Data are shown as mean ± SD with three biological replications. Statistically significant differences were determined with the pair-wise Student’s t -test, and significance levels were denoted by *** (p<0.001). Different letters above the bars indicated statistically significant differences (p<0.05), as determined by ANOVA followed with a Turkey’s HSD multiple comparison test in SPSS 20.0 software.

Article Snippet: We therefore attempted to identify chemical compounds that either activate or inhibit CcTRPM . qRT-PCR results revealed that menthol treatment with all three concentrations significantly increased the mRNA expression of CcTRPM and antagonist (HY-112430, MedChemExpress, Shanghai, China) treatment with all three concentrations markedly reduced CcTRPM transcription ( ).

Techniques: Quantitative RT-PCR, Expressing, Imaging, Recombinant, Plasmid Preparation, Comparison, Software

Journal: eLife

Article Title: miR-252 targeting temperature receptor CcTRPM to mediate the transition from summer-form to winter-form of Cacopsylla chinensis

doi: 10.7554/eLife.88744

Figure Lengend Snippet: ( A ) The putative miR-252 binding sites in CcTRPM 3’UTR were predicted by miRanda and Targetscan. ( B ) The expression profiles of miR-252 in response to different temperatures of 25°C and 10°C by qRT-PCR. ( C ) In vitro validation of the target interactions between miR-252 and CcTRPM by dual luciferase reporter assays. ( D ) In vivo demonstration of miR-252 targeting CcTRPM by RNA-binding protein immunoprecipitation (RIP) assay. ( E ) Representative confocal images of miR-252 and CcTRPM in 1st, 3rd and 5th instar of SF. Scale bar is 0.5 mm. The signals of DAPI and CcTRPM -Cy3 are same as the above describing. miR-252-FAM: the miR-252 signal was labeled with FAM and visualized in green. Merge: merged imaging of co-localization of cell nucleus, CcTRPM and miR-252. ( F ) Effect of miR-252 agomir and antagomir treatment 3 days on the expression of CcTRPM at mRNA level. The data in 3B and 3 F are shown as the mean ± SD with three independent biological replications of at least 30 nymphs for each biological replication. Data in 3 C are presented as mean ± SD with nine biological replications. Data in 3D are presented as mean ± SD with two biological replications of three technical replicates for each biological replication. Statistically significant differences were determined with the pair-wise Student’s t -test, and significance levels were denoted by *** (p<0.001). Different letters above the bars indicated statistically significant differences (p<0.05), as determined by ANOVA followed with a Turkey’s HSD multiple comparison test in SPSS 20.0 software.

Article Snippet: We therefore attempted to identify chemical compounds that either activate or inhibit CcTRPM . qRT-PCR results revealed that menthol treatment with all three concentrations significantly increased the mRNA expression of CcTRPM and antagonist (HY-112430, MedChemExpress, Shanghai, China) treatment with all three concentrations markedly reduced CcTRPM transcription ( ).

Techniques: Binding Assay, Expressing, Quantitative RT-PCR, In Vitro, Biomarker Discovery, Luciferase, In Vivo, RNA Binding Assay, Immunoprecipitation, Labeling, Imaging, Comparison, Software

Journal: eLife

Article Title: miR-252 targeting temperature receptor CcTRPM to mediate the transition from summer-form to winter-form of Cacopsylla chinensis

doi: 10.7554/eLife.88744

Figure Lengend Snippet: ( A ) Effect of miR-252 agomir treatment on the mRNA expression of miR-252 after 3 days and 6 days under 10 °C. agomir-NC treatment was used as the control. ( B ) The mRNA expression of miR-252 after dsCcTRPM treatment 3 days and 6 days compare with the dsEGFP treatment under 10 °C. ( C–F ) Comparison of the nymph cuticle thickness, cuticle ultrastructure, cuticle chitin staining with WGA-FITC, and chitin content of SF 1st instar after treatment with agomir-NC and agomir-252 at 15 days. Scale bar in (D) is 1μm and in (E) is 100 μm. The two-way arrow indicated the cuticle thickness. The DAPI and WAG-FITC signals were same as the above describing. ( G ) The transition percent of SF 1st instar nymphs treated with agomir-NC, agomir-252, dsEGFP, dsCcTRPM, and dsCcTRPM +antagomir-252 at 15 day under 10 °C. ( H ) The phenotypes of SF 1st instar nymphs treated with agomir-NC, agomir-252, dsCcTRPM, and dsCcTRPM +antagomir-252 at 15 day under 10 °C. Scale bar is 0.5 mm. The data in 4A and 4B are shown as the mean ± SD with three independent biological replications of at least 30 nymphs for each biological replication. Data in 4C and 4F are presented as mean ± SD with three biological replications of three technical replications for each biological replication. Data in 4G are presented as mean ± SD with nine biological replications. Statistically significant differences were determined with the pair-wise Student’s t -test, and significance levels were denoted by *** (p<0.001). Different letters above the bars indicated statistically significant differences (p<0.05), as determined by ANOVA followed with a Turkey’s HSD multiple comparison test in SPSS 20.0 software.

Article Snippet: We therefore attempted to identify chemical compounds that either activate or inhibit CcTRPM . qRT-PCR results revealed that menthol treatment with all three concentrations significantly increased the mRNA expression of CcTRPM and antagonist (HY-112430, MedChemExpress, Shanghai, China) treatment with all three concentrations markedly reduced CcTRPM transcription ( ).

Techniques: Expressing, Control, Comparison, Staining, Software

Journal: eLife

Article Title: miR-252 targeting temperature receptor CcTRPM to mediate the transition from summer-form to winter-form of Cacopsylla chinensis

doi: 10.7554/eLife.88744

Figure Lengend Snippet: ( A–B ) Melting curve for qRT-PCR primers of U6 and miR-252. ( C ) Effect of agomir-252 treatment 2 day and 4 day on the mRNA expression of CcTRPM under 25 °C condition. ( D ) The phenotype of SF 1st instar nymphs treated with agomir-NC and agomir-252 at 10 day at 25 °C. Scale bar is 0.5 mm. Data are shown as the mean ± SD with three independent biological replications of at least 30 nymphs for each biological replication. Statistically significant differences were determined with the pair-wise Student’s t -test, and significance levels were denoted by *** (p<0.001).

Article Snippet: We therefore attempted to identify chemical compounds that either activate or inhibit CcTRPM . qRT-PCR results revealed that menthol treatment with all three concentrations significantly increased the mRNA expression of CcTRPM and antagonist (HY-112430, MedChemExpress, Shanghai, China) treatment with all three concentrations markedly reduced CcTRPM transcription ( ).

Techniques: Quantitative RT-PCR, Expressing

Journal: eLife

Article Title: miR-252 targeting temperature receptor CcTRPM to mediate the transition from summer-form to winter-form of Cacopsylla chinensis

doi: 10.7554/eLife.88744

Figure Lengend Snippet: ( A ) Diagram of de novo biosynthesis of insect chitin. Tre: trehalase. HK: hexokinase. G6PI: glucose-6-phosphate isomerase. GFAT: fructose-6-phosphate aminotransferase. GNA: glucosamine-6-phosphate acetyltransferase. AGM: N-acetylglucosamine phosphate mutase. UAP: UDP- N-acetylglucosamine pyrophosphorylase. CHS: chitin synthase. ( B ) A heat map was constructed from the expression levels of chitin biosynthesis enzyme transcripts in C. chinensis after 25°C and 10°C treatment at 3 d, 6 d, and 10 d. ( C ) Effect of RNAi knockdown of 14 transcripts covering all the enzymes in the chitin biosynthesis pathway on the transition percent of SF 1st instar nymphs under 10 °C. ( D–F ) Comparison of the nymph cuticle thickness, cuticle chitin content, and cuticle chitin staining with WGA-FITC of SF 1st instar after treatment with dsEGFP, dsCcTre1, and dsCcCHS1 at 15 d. The WAG-FITC signal was same as the above describing. Scale bar in (F) is 100 μm and 0.5 mm, respectively. ( G–H ) Effect of dsCcTRPM and agomir-252 treatments on the mRNA expression of CcTre1 at 3 day and 6 day under 10 °C. ( I–J ) Effect of dsCcTRPM and agomir-252 treatments on the mRNA expression of CcCHS1 at 3 day and 6 day under 10 °C. The data in ( C ) are shown as the mean ± SD with six independent biological replications of at least 30 nymphs for each biological replication. Data in ( D and E ) are presented as mean ± SD with three biological replications of three technical replications for each biological replication. Data in ( G-J ) are presented as mean ± SD with three biological replications. Statistically significant differences were determined with the pair-wise Student’s t -test, and significance levels were denoted by * (p<0.05), ** (p<0.01), and *** (p<0.001).

Article Snippet: We therefore attempted to identify chemical compounds that either activate or inhibit CcTRPM . qRT-PCR results revealed that menthol treatment with all three concentrations significantly increased the mRNA expression of CcTRPM and antagonist (HY-112430, MedChemExpress, Shanghai, China) treatment with all three concentrations markedly reduced CcTRPM transcription ( ).

Techniques: Construct, Expressing, Knockdown, Comparison, Staining

Journal: CNS Neuroscience & Therapeutics

Article Title: GPR30‐mediated estrogenic regulation of actin polymerization and spatial memory involves SRC‐1 and PI3K‐mTORC2 in the hippocampus of female mice

doi: 10.1111/cns.13108

Figure Lengend Snippet: Selective activation of mTORC2 reversed G15‐induced spatial memory disorder and actin depolymerization. The sham animals were in diestrous cycling (evidenced by vaginal smear). The mice injected with DMSO were used as the control. A, Flowchart of the experiments. B, A4 reversed G15‐induced learning impairment from days 3 to day 5. C, The swimming tracks of mice in each quadrant. D, The time spent in the target quadrant of animals. E‐G, The total distance traveled and swimming speed. H, A4 rescued G15‐induced downregulation of p‐AKT. I, A4 reversed G15‐induced decrease in F‐actin/G‐actin ratio. A4: mTORC2 activator A‐443654. Data are shown as the mean ± SEM. ** P < 0.01 compared with other groups (repeated measures of two‐way ANOVA, LSD test for water maze test and one‐way ANOVA, LSD test for Western blot analysis)

Article Snippet: The mTORC2‐specific activator A4 (HY‐10425; Medchem Express, Shanghai, China) was prepared with DMSO and intraperitoneally injected with a dose of 2.5 mg/kg body weight as previous reports., Animals in the DMSO and/or sham groups received an injection of an equal amount of vehicle (60% DMSO + 40% sterile saline solution) as indicated in each experiment.

Techniques: Activation Assay, Injection, Control, Western Blot