Journal: International journal of molecular sciences

Article Title: Expression of the Calcitonin Receptor-like Receptor (CALCRL) in Normal and Neoplastic Tissues.

doi: 10.3390/ijms24043960

Figure Lengend Snippet: Figure 3. Double-labelling immunocytochemical analysis of calcitonin receptor-like receptor (CAL- CRL) expression (A,D,G) and the expression of receptor activity-modifying protein (RAMP) 1 (B), RAMP2 (E), or RAMP3 (H) in BON-1 cells. Labelling of CALCRL was visualised using Cy3-conjugated anti-rabbit antibody (red). Labelling of RAMP1, RAMP2, or RAMP3 was visualised using Alexa Fluor 488-conjugated rabbit anti-RAMP1, -RAMP2, or -RAMP3 antibody (green). Overlapping expression is represented by orange/yellow colour (C,F,I). Blue colour represents 4′,6-diamidino-2-phenylindole (DAPI)-stained DNA. Scale bar: 100 µm.

Article Snippet: The cells were then washed thoroughly with phosphate-buffered saline and incubated overnight at 4 ◦C with Alexa 488-conjugated rabbit polyclonal anti-RAMP1, RAMP2, or RAMP3 antibody (1:100 dilution; Bioss Antibodies, Woburn, MA, USA; catalogue numbers, bs-1567R-A488; bs-11971R-A488; bs-11972R-A488; the antibodies were tested beforehand for specificity by means of siRNA knockdown experiments in BON-1 cells, which (as could also be shown in the present paper) endogenously express all three RAMP proteins [siRNAs used: RAMP1, sc-40894; RAMP2, sc-3678; RAMP3, sc-40896; Santa Cruz Biotechnology; Supplemental Figure S7] and by means of peptide neutralisations in RAMPpositive tissues (duodenum; placenta); Supplemental Figure S8).

Techniques: Expressing, Activity Assay, Staining

Journal: International journal of molecular sciences

Article Title: Expression of the Calcitonin Receptor-like Receptor (CALCRL) in Normal and Neoplastic Tissues.

doi: 10.3390/ijms24043960

Figure Lengend Snippet: Figure 6. Double-labelling immunohistochemical analysis of calcitonin receptor-like receptor (CAL- CRL) expression (A,E,I) and the expression of receptor activity-modifying protein (RAMP) 1 (B), RAMP2 (F), or RAMP3 (J) in human pituitary tissue. Labelling of CALCRL was visualised using Cy3-conjugated anti-rabbit antibody (red). Labelling of RAMP1, RAMP2, or RAMP3 was visualised using Alexa Fluor 488-conjugated rabbit anti-RAMP1, -RAMP2, or -RAMP3 antibody (green). Over- lapping expression is represented by orange/yellow colour (C,D,G,H,K,L). Blue colour represents 4′,6-diamidino-2-phenylindole (DAPI)-stained DNA. Arrows: CALCRL-negative cells that express RAMP1 or RAMP3. (D,H,L) represent enlarged sections of (C,G,K). Scale bar: 100 µm (A–C,E–G,I–K); 50 µm (D,H,L).

Article Snippet: The cells were then washed thoroughly with phosphate-buffered saline and incubated overnight at 4 ◦C with Alexa 488-conjugated rabbit polyclonal anti-RAMP1, RAMP2, or RAMP3 antibody (1:100 dilution; Bioss Antibodies, Woburn, MA, USA; catalogue numbers, bs-1567R-A488; bs-11971R-A488; bs-11972R-A488; the antibodies were tested beforehand for specificity by means of siRNA knockdown experiments in BON-1 cells, which (as could also be shown in the present paper) endogenously express all three RAMP proteins [siRNAs used: RAMP1, sc-40894; RAMP2, sc-3678; RAMP3, sc-40896; Santa Cruz Biotechnology; Supplemental Figure S7] and by means of peptide neutralisations in RAMPpositive tissues (duodenum; placenta); Supplemental Figure S8).

Techniques: Immunohistochemical staining, Expressing, Activity Assay, Staining

Journal: International journal of molecular sciences

Article Title: Expression of the Calcitonin Receptor-like Receptor (CALCRL) in Normal and Neoplastic Tissues.

doi: 10.3390/ijms24043960

Figure Lengend Snippet: Figure 7. Double-labelling immunohistochemical analysis of calcitonin receptor-like receptor (CAL- CRL) expression (A,E,I) and the expression of receptor activity-modifying protein (RAMP) 1 (B), RAMP2 (F), or RAMP3 (J) in human duodenal tissue. Labelling of CALCRL was visualised using Cy3-conjugated anti-rabbit antibody (red). Labelling of RAMP1, RAMP2, or RAMP3 was visu- alised using Alexa Fluor 488-conjugated rabbit anti-RAMP1, -RAMP2, or -RAMP3 antibody (green). Overlapping expression is represented by orange/yellow colour (C,D,G,H,K,L). Blue colour repre- sents 4′,6-diamidino-2-phenylindole (DAPI)-stained DNA. Arrows: Neuroendocrine cells. (D,H,L) represent enlarged sections of (C,G,K). Scale bar: 100 µm (A–C,E–G,I–K), 50 µm (D,H,L).

Article Snippet: The cells were then washed thoroughly with phosphate-buffered saline and incubated overnight at 4 ◦C with Alexa 488-conjugated rabbit polyclonal anti-RAMP1, RAMP2, or RAMP3 antibody (1:100 dilution; Bioss Antibodies, Woburn, MA, USA; catalogue numbers, bs-1567R-A488; bs-11971R-A488; bs-11972R-A488; the antibodies were tested beforehand for specificity by means of siRNA knockdown experiments in BON-1 cells, which (as could also be shown in the present paper) endogenously express all three RAMP proteins [siRNAs used: RAMP1, sc-40894; RAMP2, sc-3678; RAMP3, sc-40896; Santa Cruz Biotechnology; Supplemental Figure S7] and by means of peptide neutralisations in RAMPpositive tissues (duodenum; placenta); Supplemental Figure S8).

Techniques: Immunohistochemical staining, Expressing, Activity Assay, Staining

Journal: International journal of molecular sciences

Article Title: Expression of the Calcitonin Receptor-like Receptor (CALCRL) in Normal and Neoplastic Tissues.

doi: 10.3390/ijms24043960

Figure Lengend Snippet: Figure 8. Double-labelling immunohistochemical analysis of calcitonin receptor-like receptor (CAL- CRL) expression (A,E,I) and the expression of receptor activity-modifying protein (RAMP) 1 (B), RAMP2 (F), or RAMP3 (J) in human pancreatic tissue. Labelling of CALCRL was visualised using Cy3-conjugated anti-rabbit antibody (red). Labelling of RAMP1, RAMP2, or RAMP3 was visualised using Alexa Fluor 488-conjugated rabbit anti-RAMP1, -RAMP2, or -RAMP3 antibody (green). Over- lapping expression is represented by orange/yellow colour (C,D,G,H,K,L). Asterisks: Pancreatic islets. Blue colour represents 4′,6-diamidino-2-phenylindole (DAPI)-stained DNA. (D,H,L) represent enlarged sections of (C,G,K). Scale bar: 100 µm (A–C,E–G,I–K), 50 µm (D,H,L).

Article Snippet: The cells were then washed thoroughly with phosphate-buffered saline and incubated overnight at 4 ◦C with Alexa 488-conjugated rabbit polyclonal anti-RAMP1, RAMP2, or RAMP3 antibody (1:100 dilution; Bioss Antibodies, Woburn, MA, USA; catalogue numbers, bs-1567R-A488; bs-11971R-A488; bs-11972R-A488; the antibodies were tested beforehand for specificity by means of siRNA knockdown experiments in BON-1 cells, which (as could also be shown in the present paper) endogenously express all three RAMP proteins [siRNAs used: RAMP1, sc-40894; RAMP2, sc-3678; RAMP3, sc-40896; Santa Cruz Biotechnology; Supplemental Figure S7] and by means of peptide neutralisations in RAMPpositive tissues (duodenum; placenta); Supplemental Figure S8).

Techniques: Immunohistochemical staining, Expressing, Activity Assay, Staining

Journal: International journal of molecular sciences

Article Title: Expression of the Calcitonin Receptor-like Receptor (CALCRL) in Normal and Neoplastic Tissues.

doi: 10.3390/ijms24043960

Figure Lengend Snippet: Figure 9. Double-labelling immunohistochemical analysis of calcitonin receptor-like receptor (CAL- CRL) expression (A,E,I) and the expression of receptor activity-modifying protein (RAMP) 1 (B), RAMP2 (F), or RAMP3 (J) in human adrenocortical tissue. Labelling of CALCRL was visualised using Cy3-conjugated anti-rabbit antibody (red). Labelling of RAMP1, RAMP2, or RAMP3 was visualised using Alexa Fluor 488-conjugated rabbit anti-RAMP1, -RAMP2, or -RAMP3 antibody (green). Overlapping expression is represented by orange/yellow colour (C,D,G,H,K,L). Blue colour represents 4′,6-diamidino-2-phenylindole (DAPI)-stained DNA. (D,H,L) represent enlarged sections of (C,G,K). Scale bar: 100 µm (A–C,E–G,I–K), 50 µm (D,H,L).

Article Snippet: The cells were then washed thoroughly with phosphate-buffered saline and incubated overnight at 4 ◦C with Alexa 488-conjugated rabbit polyclonal anti-RAMP1, RAMP2, or RAMP3 antibody (1:100 dilution; Bioss Antibodies, Woburn, MA, USA; catalogue numbers, bs-1567R-A488; bs-11971R-A488; bs-11972R-A488; the antibodies were tested beforehand for specificity by means of siRNA knockdown experiments in BON-1 cells, which (as could also be shown in the present paper) endogenously express all three RAMP proteins [siRNAs used: RAMP1, sc-40894; RAMP2, sc-3678; RAMP3, sc-40896; Santa Cruz Biotechnology; Supplemental Figure S7] and by means of peptide neutralisations in RAMPpositive tissues (duodenum; placenta); Supplemental Figure S8).

Techniques: Immunohistochemical staining, Expressing, Activity Assay, Staining

Journal: International journal of molecular sciences

Article Title: Expression of the Calcitonin Receptor-like Receptor (CALCRL) in Normal and Neoplastic Tissues.

doi: 10.3390/ijms24043960

Figure Lengend Snippet: Figure 10. Double-labelling immunohistochemical analysis of calcitonin receptor-like receptor (CAL- CRL) expression (A,E,I) and the expression of receptor activity-modifying protein (RAMP) 1 (B), RAMP2 (F), or RAMP3 (J) in human placental tissue. Labelling of CALCRL was visualised using Cy3-conjugated anti-rabbit antibody (red). Labelling of RAMP1, RAMP2, or RAMP3 was visualised using Alexa Fluor 488-conjugated rabbit anti-RAMP1, -RAMP2, or -RAMP3 antibody (green). Over- lapping expression is represented by orange/yellow colour (C,D,G,H,K,L). Blue colour represents 4′,6-diamidino-2-phenylindole (DAPI)-stained DNA. (D,H,L) represent enlarged sections of (C,G,K). Arrows: Syncytiotrophoblast cells; arrowheads: Capillary endothelia; asterisks: Non-specifically stained erythrocytes. Scale bar: 100 µm (A–C,E–G,I–K), 50 µm (D,H,L).

Article Snippet: The cells were then washed thoroughly with phosphate-buffered saline and incubated overnight at 4 ◦C with Alexa 488-conjugated rabbit polyclonal anti-RAMP1, RAMP2, or RAMP3 antibody (1:100 dilution; Bioss Antibodies, Woburn, MA, USA; catalogue numbers, bs-1567R-A488; bs-11971R-A488; bs-11972R-A488; the antibodies were tested beforehand for specificity by means of siRNA knockdown experiments in BON-1 cells, which (as could also be shown in the present paper) endogenously express all three RAMP proteins [siRNAs used: RAMP1, sc-40894; RAMP2, sc-3678; RAMP3, sc-40896; Santa Cruz Biotechnology; Supplemental Figure S7] and by means of peptide neutralisations in RAMPpositive tissues (duodenum; placenta); Supplemental Figure S8).

Techniques: Immunohistochemical staining, Expressing, Activity Assay, Staining

Journal: International journal of molecular sciences

Article Title: Expression of the Calcitonin Receptor-like Receptor (CALCRL) in Normal and Neoplastic Tissues.

doi: 10.3390/ijms24043960

Figure Lengend Snippet: Figure 12. Double-labelling immunohistochemical analysis of calcitonin receptor-like receptor (CAL- CRL) expression (A,E,I) and the expression of receptor activity-modifying protein (RAMP) 1 (B), RAMP2 (F), or RAMP3 (J) in medullary thyroid cancer tissue. Labelling of CALCRL was visualised using Cy3-conjugated anti-rabbit antibody (red). Labelling of RAMP1, RAMP2, or RAMP3 was visualised using Alexa Fluor 488-conjugated rabbit anti-RAMP1, -RAMP2, or -RAMP3 antibody (green). Overlapping expression is represented by orange/yellow colour. Blue colour represents 4′,6- diamidino-2-phenylindole (DAPI)-stained DNA. (D,H,L) represent (C,D,G,H,K,L) enlarged sections of (C,G,K). Scale bar: 100 µm (A–C,E–G,I–K), 50 µm (D,H,L).

Article Snippet: The cells were then washed thoroughly with phosphate-buffered saline and incubated overnight at 4 ◦C with Alexa 488-conjugated rabbit polyclonal anti-RAMP1, RAMP2, or RAMP3 antibody (1:100 dilution; Bioss Antibodies, Woburn, MA, USA; catalogue numbers, bs-1567R-A488; bs-11971R-A488; bs-11972R-A488; the antibodies were tested beforehand for specificity by means of siRNA knockdown experiments in BON-1 cells, which (as could also be shown in the present paper) endogenously express all three RAMP proteins [siRNAs used: RAMP1, sc-40894; RAMP2, sc-3678; RAMP3, sc-40896; Santa Cruz Biotechnology; Supplemental Figure S7] and by means of peptide neutralisations in RAMPpositive tissues (duodenum; placenta); Supplemental Figure S8).

Techniques: Immunohistochemical staining, Expressing, Activity Assay, Staining

Journal: International journal of molecular sciences

Article Title: Expression of the Calcitonin Receptor-like Receptor (CALCRL) in Normal and Neoplastic Tissues.

doi: 10.3390/ijms24043960

Figure Lengend Snippet: Figure 13. Double-labelling immunohistochemical analysis of calcitonin receptor-like receptor (CAL- CRL) expression (A,E,I) and the expression of receptor activity-modifying protein (RAMP) 1 (B), RAMP2 (F), or RAMP3 (J) in renal clear cell cancer tissue. Labelling of CALCRL was visualised using Cy3-conjugated anti-rabbit antibody (red). Labelling of RAMP1, RAMP2, or RAMP3 was visualised using Alexa Fluor 488-conjugated rabbit anti-RAMP1, -RAMP2, or -RAMP3 antibody (green). Overlapping expression is represented by orange/yellow colour (C,D,G,H,K,L). Blue colour represents 4′,6-diamidino-2-phenylindole (DAPI)-stained DNA. (D,H,L) represent enlarged sections of (C,G,K). Scale bar: 100 µm (A–C,E–G,I–K), 50 µm (D,H,L).

Article Snippet: The cells were then washed thoroughly with phosphate-buffered saline and incubated overnight at 4 ◦C with Alexa 488-conjugated rabbit polyclonal anti-RAMP1, RAMP2, or RAMP3 antibody (1:100 dilution; Bioss Antibodies, Woburn, MA, USA; catalogue numbers, bs-1567R-A488; bs-11971R-A488; bs-11972R-A488; the antibodies were tested beforehand for specificity by means of siRNA knockdown experiments in BON-1 cells, which (as could also be shown in the present paper) endogenously express all three RAMP proteins [siRNAs used: RAMP1, sc-40894; RAMP2, sc-3678; RAMP3, sc-40896; Santa Cruz Biotechnology; Supplemental Figure S7] and by means of peptide neutralisations in RAMPpositive tissues (duodenum; placenta); Supplemental Figure S8).

Techniques: Immunohistochemical staining, Expressing, Activity Assay, Staining