Journal: Nature cell biology

Article Title: An androgen receptor switch underlies lineage infidelity in treatment-resistant prostate cancer

doi: 10.1038/s41556-021-00743-5

Figure Lengend Snippet: a, Heat map depicting normalized z-score for genes in the pEZH2-T350 signature in NPp53 GEMMs (Gene Expression Omnibus (GEO) accession: GSE92721). ABI, abiraterone acetate; NR, exceptional non-responder. b, pEZH2-T350 signature score in patient tumours from Aggarwal (ref. 5) (P = 0.0019) and Beltran (ref. 4) (P = 0.012). Violin plots show mean and interquartile range, with significance assessed using a two-tailed unpaired t-test. AdPC, prostate adenocarcinoma; tNEPC, treatment-induced neuroendocrine prostate cancer. c, Correlation between pEZH2-T350 and ASC scores in NEPC tumours from ref. 4. Each dot represents a patient tumour showing RB1 loss-of-function score. d, Immunohistochemical staining in patient-derived NEPC organoids. Scale bar, 100 μm. e, Immunohistochemical staining in PB-Cre4:Ptenf/f and PB-Cre4:Ptenf/f;Rb1f/f GEMMs. Scale bars: 200 μm (prostate), 1 mm (liver), 20 μm (insets). f, pEZH2-T350 score in PTEN-null GEMMs with RB1 deletion alone (P = 0.022) or concomitant with TP53 loss (P = 0074; GEO accession: GSE90891). Dots represent individual tumours. Box plot shows mean and interquartile range, with significance assessed using a two-tailed unpaired t-test. g, Volcano plot of RNA-seq from PB-Cre4:Ptenf/f and PB-Cre4:Ptenf/f;Rb1f/f GEMMs (GEO accession: GSE90891). Each dot represents a gene, with those in the ‘Reactome neuronal system’ (neuronal) and ‘Wong embryonic stem cell core’ (plasticity) MSigDB pathways highlighted. h, Immunoblot in 16DCRPC cells with stable RB1 knockdown treated with CDK1 inhibitor (5 μM RO-3306, 6 h). shNS, non-silencing control. i, Heat map of gene expression (by rtPCR) in CRPCcrEZH2 cells with shRB1 expressing EZH2WT, EZH2T350A or EZH2T350D mutant constructs following treatment with DMSO or ENZ (10 μM) for 7 d. Expression is reported relative to parental cells (n = 3). j, Left: quantification of spheroids in CRPCcrEZH2 cells with shRB1 expressing EZH2T350A or EZH2T350D treated with DMSO or ENZ (10 μM). Data reported as mean ± s.d., with significance evaluated at the end point (P = 0.003, two-tailed unpaired t-test; n = 3). Right: images at 5 d after ENZ treatment. Scale bar, 100 μm. k, Left: neurite length measured using IncuCyte in CRPCcrEZH2 cells with shRB1 expressing EZH2T350A or EZH2T350D and treated with DMSO or ENZ (10 μM). Right: phase-contrast images at 96 h after ENZ treatment, with neurite-like extensions highlighted by NeuroTrack. Data reported as mean ± s.d. with significance evaluated at end-point (P = 0.0003, two-tailed unpaired t-test; n = 3). Scale bar, 100 μm.

Article Snippet: The shRB1 (#25641), pcDNA3-EZH2 S21A (#42663), pcDNA3-EZH2 S21D (#42664) and control (#8453) plasmids were available from Addgene.

Techniques: Expressing, Two Tailed Test, Immunohistochemical staining, Staining, Derivative Assay, RNA Sequencing Assay, Western Blot, Reverse Transcription Polymerase Chain Reaction, Mutagenesis, Construct

Journal: Microscopy and Microanalysis

Article Title: LPS- and LTA-Induced Expression of TLR4, MyD88, and TNF-α in Lymph Nodes of the Akkaraman and Romanov Lambs

doi: 10.1017/s1431927622012314

Figure Lengend Snippet: Fig. 3. TLR4, MyD88, and TNF-α immunoreactivity in the bronchial lymph nodes of Akkaraman lambs in LTA, LPS, LTA + LPS, control groups and negative control (e,j,o). K, capsule; T, trabeculae; LF, lymph follicle; V, blood vessel; Sg, centrum germinativum; Sc, subcapsuler sinus; Mc, medullary cord; arrows, TLR4 immuno- positive smooth muscle cells; arrow heads, TNF-α immunopositive immune cells. Scale bars, 20 μm (a–d,f–i,l–n) and 50 μm (e,j,k,o).

Article Snippet: The sections were incubated at 4 °C overnight with mouse monoclonal TLR4 (Novus Biologicals, CO, USA, NB100-56566, 1:100 dilution), rabbit polyclonal MyD88 (Novus Biologicals, CO, USA, NB100-56598, 1:100 dilution), and mouse monoclonal TNF-α primary antibodies (Bio-Rad Laboratories MCA905GA, 1:75 dilution).

Techniques: Control, Negative Control

Journal: Microscopy and Microanalysis

Article Title: LPS- and LTA-Induced Expression of TLR4, MyD88, and TNF-α in Lymph Nodes of the Akkaraman and Romanov Lambs

doi: 10.1017/s1431927622012314

Figure Lengend Snippet: Fig. 4. TLR4, MyD88, and TNF-α immunoreactivity in the bronchial lymph nodes of Romanov lambs in LTA, LPS, LTA + LPS, control groups, and negative control (e,j, o). K, capsule; T, trabeculae; LF, lymph follicle; V, blood vessel; Mc, medullary cord; Ms, medullary sinus; arrows, TLR4 immunopositive smooth muscle cells; arrow heads, TNF-α immunopositive immune cells. Scale bars, 20 μm (a–d,g–i,l–n) and 50 μm (e,f,j,k,o).

Article Snippet: The sections were incubated at 4 °C overnight with mouse monoclonal TLR4 (Novus Biologicals, CO, USA, NB100-56566, 1:100 dilution), rabbit polyclonal MyD88 (Novus Biologicals, CO, USA, NB100-56598, 1:100 dilution), and mouse monoclonal TNF-α primary antibodies (Bio-Rad Laboratories MCA905GA, 1:75 dilution).

Techniques: Control, Negative Control

Journal: Microscopy and Microanalysis

Article Title: LPS- and LTA-Induced Expression of TLR4, MyD88, and TNF-α in Lymph Nodes of the Akkaraman and Romanov Lambs

doi: 10.1017/s1431927622012314

Figure Lengend Snippet: Fig. 5. TLR4, MyD88, and TNF-α immunoreactivity in the mandibular lymph nodes of Akkaraman lambs in LTA, LPS, LTA + LPS, control groups, and negative control (e,j,o). K, capsule; T, trabeculae; LF, lymph follicle; V, blood vessel; Sc, subcapsuler sinus; Ms, medullary sinus; Mc, medullary cord; arrows, smooth muscle cells; arrow heads, TNF-α immunopositive immune cells. Scale bars, 20 μm (a–d,f–i,k–n) and 50 μm (e,j,o).

Article Snippet: The sections were incubated at 4 °C overnight with mouse monoclonal TLR4 (Novus Biologicals, CO, USA, NB100-56566, 1:100 dilution), rabbit polyclonal MyD88 (Novus Biologicals, CO, USA, NB100-56598, 1:100 dilution), and mouse monoclonal TNF-α primary antibodies (Bio-Rad Laboratories MCA905GA, 1:75 dilution).

Techniques: Control, Negative Control

Journal: Microscopy and Microanalysis

Article Title: LPS- and LTA-Induced Expression of TLR4, MyD88, and TNF-α in Lymph Nodes of the Akkaraman and Romanov Lambs

doi: 10.1017/s1431927622012314

Figure Lengend Snippet: Fig. 6. TLR4, MyD88, and TNF-α immunoreactivity in the mandibular lymph nodes of Romanov lambs in LTA, LPS, LTA + LPS, control groups, and negative control (e,j,o). K, capsule; T, trabeculae; LF, lymph follicle; V, blood vessel; Sg, centrum germinativum; arrows, TLR4 immunopositive smooth muscle cells. Scale bars, 20 μm (a–d,g–i,l–n) and 50 μm (e,f,j,k,o).

Article Snippet: The sections were incubated at 4 °C overnight with mouse monoclonal TLR4 (Novus Biologicals, CO, USA, NB100-56566, 1:100 dilution), rabbit polyclonal MyD88 (Novus Biologicals, CO, USA, NB100-56598, 1:100 dilution), and mouse monoclonal TNF-α primary antibodies (Bio-Rad Laboratories MCA905GA, 1:75 dilution).

Techniques: Control, Negative Control

Journal: Microscopy and Microanalysis

Article Title: LPS- and LTA-Induced Expression of TLR4, MyD88, and TNF-α in Lymph Nodes of the Akkaraman and Romanov Lambs

doi: 10.1017/s1431927622012314

Figure Lengend Snippet: Fig. 7. Western blot analysis of TLR4, MyD88, and TNF-α proteins in the bronchial (a–c) and mandibular (d–f) lymph nodes of Akkaraman and Romanov lambs. Bronchial and mandibular lymph node tissues were obtained from LTA, LPS, LTA + LPS, and control groups (c). Expressions of TLR4, MyD88, and TNF-α molecules were assessed by Western blot as described in the Method section. The graphs below the representative blots show densitometric quantitation of protein bands. Beta actin was used as an internal loading control protein. Data are the mean ± SEM of three replicates in each group.

Article Snippet: The sections were incubated at 4 °C overnight with mouse monoclonal TLR4 (Novus Biologicals, CO, USA, NB100-56566, 1:100 dilution), rabbit polyclonal MyD88 (Novus Biologicals, CO, USA, NB100-56598, 1:100 dilution), and mouse monoclonal TNF-α primary antibodies (Bio-Rad Laboratories MCA905GA, 1:75 dilution).

Techniques: Western Blot, Control, Quantitation Assay

Journal: Microscopy and Microanalysis

Article Title: LPS- and LTA-Induced Expression of TLR4, MyD88, and TNF-α in Lymph Nodes of the Akkaraman and Romanov Lambs

doi: 10.1017/s1431927622012314

Figure Lengend Snippet: Fig. 8. Gene expression levels of TLR4, MyD88, andTNF-α in mandibular and bronchial lymph nodes of LTA, LPS, LTA + LPS, and control groups of Akkaraman and Romanov lambs.

Article Snippet: The sections were incubated at 4 °C overnight with mouse monoclonal TLR4 (Novus Biologicals, CO, USA, NB100-56566, 1:100 dilution), rabbit polyclonal MyD88 (Novus Biologicals, CO, USA, NB100-56598, 1:100 dilution), and mouse monoclonal TNF-α primary antibodies (Bio-Rad Laboratories MCA905GA, 1:75 dilution).

Techniques: Gene Expression, Control