Journal: International Journal of Molecular Sciences

Article Title: Tandem Expression of a Mobile RNA and Its RNA-Binding Protein(s) Enhances Tuber Productivity in Potato

doi: 10.3390/ijms242115754

Figure Lengend Snippet: Evaluation of earliness for tuberization in Désirée CC lines. ( A ) Image of wild-type (WT) and CC lines of S. tuberosum cv. Désirée growing in the greenhouse. ( B ) Representative images of WT and CC Désirée lines after 12 weeks of transfer to soil. Lower yellow leaves in CC lines indicate their earliness for maturity. The enhanced expression of StCDF1 in these lines (shown later in A) could be the reason. Scale bar = 5 cm. ( C ) In vitro tuber induction experiment (8% sucrose) showed an earliness in transgenic CC Désirée lines compared to WT. ( D ) Average number of stolons per plant after 3-week of transfer to greenhouse. Student’s t -test was used at p < 0.05 (*** p < 0.001, **** p < 0.0001, ns, not significant). Mean values ±SEM are represented in the graph. ( E ) Evaluation of earliness for tuberization in greenhouse after 3-weeks. The graph represents the percentage of plants that formed stolons and/or swollen stolon after three weeks of growth. For panels ( C , E ), no statistical test is performed as the data is plotted as the percentage of nodes or plants tuberizing. For panels ( D , E ), values inside the bars represent the number of plants per line ( n ). #1 and #2 are the two independent lines of respective CC used throughout the studies (as mentioned in ).

Article Snippet: To evaluate the performance of Désirée CC lines under field conditions, we grew them in a contained facility at ICAR Central Potato Research Institute (CPRI), Shimla ( A–C).

Techniques: Expressing, In Vitro, Transgenic Assay

Journal: International Journal of Molecular Sciences

Article Title: Tandem Expression of a Mobile RNA and Its RNA-Binding Protein(s) Enhances Tuber Productivity in Potato

doi: 10.3390/ijms242115754

Figure Lengend Snippet: Expression of tuber marker genes and assessment of tuber productivity in Désirée CC lines under greenhouse conditions. Heat maps depict the relative expression of key tuberization genes in leaves ( A ) and stolons ( B ) of 3-week-old greenhouse plants. Data represents mean of three biological and three technical replicates. Potato EIF3e was used for normalization of gene expression and Ct values are represented next to the respective rows. #1 and #2 are the two independent lines of respective CC used throughout the studies (as mentioned in ). ( C ) Average tuber weight in transgenic Désirée CC lines from 19-week-old plants from greenhouse. The numbers below violin plots represent the number of plants per line (n). In the respective violin plot, a thick central line represents a median and the dotted lines indicate the lower and higher quartiles of the corresponding data points. In panels ( A – C ), Student’s t -test was used at p < 0.05 (* p < 0.05, ** p < 0.001, *** p < 0.005, **** p < 0.0001, ns, not significant). ( D ) Tuber images from greenhouse plants. For imaging, tubers were pooled from 15 plants for each line. Scale bar = 2 cm. The experiment for tuber productivity from greenhouse grown plants was performed two times and similar observations were found. The current data is from the second experiment. Abbreviations: BEL , BEL1-LIKE transcription factor; CDF1, CYCLING DOF FACTOR ; CO2, CONSTANS2 ; GA2OX1, GIBBERELLIN 2 OXIDASE 1 ; IT1, IDENTITY OF TUBER 1 ; PIN4, PIN FORMED 4 ; PTB1/6, POLYPYRIMIDINE TRACT-BINDING PROTEINS 1/6 ; SP5G, SELF-PRUNING 5G ; SP6A, SELF-PRUNING 6A ; SWEET11B, SUGARS WILL EVENTUALLY BE EXPORTED TRANSPORTER 11B .

Article Snippet: To evaluate the performance of Désirée CC lines under field conditions, we grew them in a contained facility at ICAR Central Potato Research Institute (CPRI), Shimla ( A–C).

Techniques: Expressing, Marker, Gene Expression, Transgenic Assay, Imaging, Binding Assay

Journal: International Journal of Molecular Sciences

Article Title: Tandem Expression of a Mobile RNA and Its RNA-Binding Protein(s) Enhances Tuber Productivity in Potato

doi: 10.3390/ijms242115754

Figure Lengend Snippet: Evaluation of tuber productivity in Désirée CC lines under field conditions. ( A ) Tuber harvest data of the transgenic Désirée CC plants grown in field conditions for 19 weeks at the ICAR Central Potato Research Institute (CPRI) located at Shimla, India. ( B ) Tuber images of transgenic control and Désirée CC lines. Scale bar = 5 cm. ( C ) Image of transgenic plants growing in the field conditions at CPRI. The field experiment was conducted two times in the year of 2021 and 2022 and similar results were obtained. #1 and #2 are the two independent lines of respective CC used throughout the studies (as mentioned in ). For panel ( A ), Student’s t -test was used at p < 0.05 (* p < 0.05, *** p < 0.001, **** p < 0.0001, ns, not significant). SEM = Standard error of mean. TC = Transgenic control. Figure contains the data obtained from the second field experiment.

Article Snippet: To evaluate the performance of Désirée CC lines under field conditions, we grew them in a contained facility at ICAR Central Potato Research Institute (CPRI), Shimla ( A–C).

Techniques: Transgenic Assay, Control

Journal: Molecular Medicine Reports

Article Title: hsa_circ_0101119 facilitates the progression of cervical cancer via an interaction with EIF4A3 to inhibit TCEAL6 expression

doi: 10.3892/mmr.2021.12293

Figure Lengend Snippet: hsa_circ_0101119 is highly expressed in CC tissues and cells. (A) Heatmap of differentially expressed circRNAs in CC tissues and normal tissues according to the online data set (GSE102686). (B) Expression level of hsa_circ_0101119 was detected via reverse transcription-quantitative PCR in CC cell lines (C-33A, SiHa, CaSki and HeLa) and normal human cervical epithelial cell line, HcerEpic. **P<0.01 vs. HcerEpic cells. (C) Expression level of hsa_circ_0101119 in CC tissues and normal tissues, according to the online data set (GSE102686). circRNA/circ, circular RNA.

Article Snippet: The four human CC cell lines (C-33A, SiHa, CaSki and HeLa) and the normal human cervical epithelial cell line, HcerEpic, were supplied by VCANBIO Cell & Gene Engineering Co., Ltd. All the cells were cultured in RPMI 1640 medium (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% (v/v) FBS (Invitrogen; Thermo Fisher Scientific, Inc.) and 1% penicillin/streptomycin (Invitrogen; Thermo Fisher Scientific, Inc.), and cultured at 37°C in a humidified 5% CO 2 incubator.

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: Molecular Medicine Reports

Article Title: hsa_circ_0101119 facilitates the progression of cervical cancer via an interaction with EIF4A3 to inhibit TCEAL6 expression

doi: 10.3892/mmr.2021.12293

Figure Lengend Snippet: hsa_circ_0101119 recruits EIF4A3 to inhibit TCEAL6 expression in CC. (A) Bioinformatics was used to predict the interaction probabilities of the RNA-binding protein EIF4A3 with hsa_circ_0101119. Predictions with probabilities >0.5 were considered ‘positive’, suggesting that the corresponding RNA and protein are likely to interact. (B) RIP assay using anti-EIF4A3 showed that EIF4A3 precipitated hsa_circ_0101119 in SiHa and HeLa cell lysates. (C) Pull down assay indicated that biotin-labeled hsa_circ_0101119 interacted with EIF4A3. (D) Bioinformatics was used to predict the interaction probabilities of EIF4A3 with TCEAL6. (E) RIP assay using anti-EIF4A3 showed that EIF4A3 precipitated TCEAL6 in SiHa and HeLa cell lysates. (F) Expression levels of EIF4A3 and TCEAL6 were detected via RT-qPCR in CC cell lines (C-33A, SiHa, CaSki and HeLa) and a normal human cervical epithelial cell line, HcerEpic. (G) Expression levels of EIF4A3 and TCEAL6 in CC tissues and normal tissues, according to the analysis of TCGA. (H) Correlation between EIF4A3 and TCEAL6 in CC samples from TCGA. (I) After transfection with sh-EIF4A3, RT-qPCR was used to detect EIF4A3 expression in SiHa and HeLa cells. (J) After transfection with sh-EIF4A3, western blotting was performed to detect the expression level of TCEAL6 in SiHa and HeLa cells. (K) After co-transfection with si-hsa_circ_0101119 and sh-EIF4A3, western blotting was performed to measure the expression level of TCEAL6 in SiHa and HeLa cells. (L) A proposed model whereby hsa_circ_0101119 sequesters EIF4A3 away from TCEAL6 mRNA, in turn suppressing TCEAL6 mRNA translation. **P<0.01 vs. IgG group (B and E); *P<0.05, **P<0.01 vs. HcerEpic cells group (F); *P<0.05 vs. normal tissues group (G); **P<0.01, vs. sh-NC group (I and J); **P<0.01 vs. sh-NC group, ## P<0.01, vs. si-hsa_circ group. (K) RIP, RNA immunoprecipitation; RT-qPCR, reverse transcription-quantitative PCR; TCGA, The Cancer Genome Atlas; sh, short hairpin RNA; NC, negative control; si, small interfering RNA; circ, circular RNA; EIF4A3, eukaryotic initiation factor 4A-3; TCEAL6, transcription elongation factor A-like 6; T, tumor; N, normal; CC, cervical cancer.

Article Snippet: The four human CC cell lines (C-33A, SiHa, CaSki and HeLa) and the normal human cervical epithelial cell line, HcerEpic, were supplied by VCANBIO Cell & Gene Engineering Co., Ltd. All the cells were cultured in RPMI 1640 medium (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% (v/v) FBS (Invitrogen; Thermo Fisher Scientific, Inc.) and 1% penicillin/streptomycin (Invitrogen; Thermo Fisher Scientific, Inc.), and cultured at 37°C in a humidified 5% CO 2 incubator.

Techniques: Expressing, RNA Binding Assay, Pull Down Assay, Labeling, Quantitative RT-PCR, Transfection, Western Blot, Cotransfection, Immunoprecipitation, Real-time Polymerase Chain Reaction, shRNA, Negative Control, Small Interfering RNA