Journal: Scientific Reports

Article Title: A novel microRNA, hsa-miR-6852 differentially regulated by Interleukin-27 induces necrosis in cervical cancer cells by downregulating the FoxM1 expression

doi: 10.1038/s41598-018-19259-4

Figure Lengend Snippet: Human Transcription factors regulated by miR-SX4 in HEK293 cells.

Article Snippet: Si-RNA against FoxM1, B-Myb, FoxO3a and Negative control siRNA were obtained from Origene.

Techniques:

Journal: Scientific Reports

Article Title: A novel microRNA, hsa-miR-6852 differentially regulated by Interleukin-27 induces necrosis in cervical cancer cells by downregulating the FoxM1 expression

doi: 10.1038/s41598-018-19259-4

Figure Lengend Snippet: miR-SX4 reduces the expression of FoxM1 target proteins. ( a ) HEK293 cells untransfected (C), Negative miRNA mimic (N) and miRNA-6852 mimic (SX4) transfected cells were harvested and protein expression levels were determined by western blot for FoxM1, regulators of FoxM1 (FoxO3a and p-FoxO3a, B-Myb and Myc) and transcriptional targets of FoxM1 (Plk1, AuroraK B, Survivin, Cdc25B, Cdc 25 C, Cdk1, Cyclin A2 and Cyclin B1). β-actin protein expression was determined for internal control. Figure 4a: i, ii, and iii images are separate western blot gel/membranes which has been stripped and reprobed with antibodies as mentioned. ( b ) HEK293 cells: C, N and SX4 transfected cells were harvested and protein expression levels were determined by western blot for upstream regulator of FoxM1, AKT/pAKT, p-p38 MAPK, ERK/p-ERK1/2, AMPKα/p-AMPKα. β-actin protein expression was determined for internal control. Figure 4b, i and iii are same western blot gel/membranes stripped and probed with another antibody; Fig. 4b, ii is another western blot gel/membrane stripped and reprobed with antibodies as mentioned. The number below the protein band represents the densitometry analysis (NIH Image J software) of protein bands normalized to β-actin or total protein levels. Bands observed for SX4 is compared with Negative miRNA mimic control (N).

Article Snippet: Si-RNA against FoxM1, B-Myb, FoxO3a and Negative control siRNA were obtained from Origene.

Techniques: Expressing, Transfection, Western Blot, Software

Journal: PLoS ONE

Article Title: Transcription Factors Involved in Prostate Gland Adaptation to Androgen Deprivation

doi: 10.1371/journal.pone.0097080

Figure Lengend Snippet: Quantitative RT-PCR determination of the mRNA content for the Mybl1 (A), Mybl2 (B) and Gata2 (C) genes in the prostate of non-castrated (NC) and castrated rats up to 7 days after surgery. The fold-change variation with respect to the controls is shown as the mean ± the standard variation (n = 3 for each time point). The asterisks indicate p<0.05. The dotted lines in each figure correspond to the fourth-power exponential fitting curve, and are shown together for the sake of direct comparison in D.

Article Snippet: RNA samples were reverse-transcribed to cDNA by a Super Script III First-Strand Kit (Invitrogen, Carlsbad, CA, USA). cDNA was mixed with TaqMan Universal PCR Master Mix (Applied Biosystems, Foster City, CA) and inventoried assays Rn01756649_g1 ( Elk-1 ), Rn01493436_m1 ( Evi-1 ), Rn01750242_m1 ( Mybl2 ), Rn01442895_m1 ( Mybl1 ), Rn01399583_m1 ( Nfkb1 ), Rn01413842_g1 ( Nfkb2 ), Rn01502266_m1 ( Rela ), Rn00583735 ( Gata2 ), Rn01648938_m1 ( Nfyb ) and Rn00573309_m1 ( Hnf4a ).

Techniques: Quantitative RT-PCR, Comparison

Journal: PLoS ONE

Article Title: Transcription Factors Involved in Prostate Gland Adaptation to Androgen Deprivation

doi: 10.1371/journal.pone.0097080

Figure Lengend Snippet: Immunohistochemical localization of selected TF (red) as indicated in the prostate of non-castrated controls (NC) and castrated rats 3 days after surgery (CAS). MYBL2 (A,B), GATA2 (C,D), EVI1 (E,F), ELK1 (G,H), NFYB (I,J), NFKB1 (K,L), NFKB2 (M,N), REL (O,P), RELA (Q,R), RELB (S,T). Nuclei were stained with DAPI (blue). White arrows indicate smooth-muscle cells. Yellow arrowheads indicate cells showing nuclear location of the TF. L = gland lumen; S = stroma. Scale bars = 50 µm.

Article Snippet: RNA samples were reverse-transcribed to cDNA by a Super Script III First-Strand Kit (Invitrogen, Carlsbad, CA, USA). cDNA was mixed with TaqMan Universal PCR Master Mix (Applied Biosystems, Foster City, CA) and inventoried assays Rn01756649_g1 ( Elk-1 ), Rn01493436_m1 ( Evi-1 ), Rn01750242_m1 ( Mybl2 ), Rn01442895_m1 ( Mybl1 ), Rn01399583_m1 ( Nfkb1 ), Rn01413842_g1 ( Nfkb2 ), Rn01502266_m1 ( Rela ), Rn00583735 ( Gata2 ), Rn01648938_m1 ( Nfyb ) and Rn00573309_m1 ( Hnf4a ).

Techniques: Immunohistochemical staining, Staining

Journal: Nature cell biology

Article Title: LRRC31 inhibits DNA repair and sensitizes breast cancer brain metastasis to radiation therapy

doi: 10.1038/s41556-020-00586-6

Figure Lengend Snippet: a, Schematic of the CRISPR screen. b, Characterization of the proliferation of control or LRRC31-knockout 231BR cells with and without irradiation (6 Gy). c, Schematic diagram of characterization of LRRC31 in mice bearing intracranial 231BR tumors. Cells were engineered to express both luciferase and GFP. d, Changes of tumor volume versus time in mice received subcutaneous inoculation of control or LRRC31-knockout 231BR cells and treated with irradiation (10 Gy). e,f, qRT-PCR analysis of the expression of miR-4796 and miR-1287 in 231Br cells transduced with lentiviral vectors for expression of the candidate miRNAs or control vector. g,h, WB analysis of the expression of KATNA and MYBL2 in 231Br cells transduced with control vector or vectors for overexpression of the indicated gene. Blot is representative of two biologically independent experiments, with similar results obtained. Unprocessed immunoblots are shown in . i-l, Clonogenic analysis of 231Br cells engineered for overexpression of miR4796 (i), miR1287 (j), KATNA1 (g) and MYBL2 (h) 7 days after irradiation. m, Characterization of the proliferation of control or LRRC31-overexpressed 231BR cells with and without irradiation (6 Gy). n, Changes of tumor volume versus time in mice received subcutaneous inoculation of control or LRRC31-overexpressed 231BR cells and treated with irradiation (10 Gy). For b, e, f, i-l, and m, data show the mean ± s.d. (n=3 biologically independent experiments). For d and n, data show the mean ± s.d. (n=3 animals). Statistical analysis was performed using the two-tailed, unpaired Student’s t-test.

Article Snippet: For overexpression of miR-1287 or miR-4796, pre-miR-1287 or pre-miR-4796 coding region was cloned into GV217 lentiviral vector (Genechem). cDNA for KATNA1 (Catalog #: MHS6278–202802607) and LRRC31 (Catalog #: MHS6278–213244531) were purchased from GE Dharmacon. cDNA of MYBL2 (Catalog #: HG14536-G) was purchased from Sino Biological.

Techniques: CRISPR, Knock-Out, Irradiation, Luciferase, Quantitative RT-PCR, Expressing, Transduction, Plasmid Preparation, Over Expression, Western Blot, Two Tailed Test