Journal: Bioengineered

Article Title: Long non-coding RNA LINC00997 silencing inhibits the progression and metastasis of colorectal cancer by sponging miR-512-3p.

doi: 10.1080/21655979.2021.1882164

Figure Lengend Snippet: Figure 3. LINC00997 silencing suppressed the migration and invasion of CRC cells. (a) LINC00997 level in several CRC cell lines was detected using RT-qPCR. **P < 0.01, ***P < 0.001 vs. HIEC. (b) The level of LINC00997 was tested with RT-qPCR after transfection with shRNA-LINC00997 into HCT116 cells. **P < 0.01, ***P < 0.001. (c and d) Cell migration was tested using wound healing assay. (e and f) The invasive ability of HCT116 cells was determined with Transwell assay. (g) Western blot analysis was used to examine the expression of MMP2 and MMP7. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Several human CRC cell lines (SW480, SW1417, SW620, Caco2, LS174T, HCT15, LoVo, HT29 and HCT116) and the normal intestinal epithelial cell line (HIEC) used in this study were obtained from American Type Culture Collection (Manassas, VA, USA).

Techniques: Migration, Quantitative RT-PCR, Transfection, shRNA, Wound Healing Assay, Transwell Assay, Western Blot, Expressing

Journal: International journal of cancer

Article Title: Identification of truncated chemokine receptor 7 in human colorectal cancer unable to localize to the cell surface and unreactive to external ligands.

doi: 10.1002/ijc.23704

Figure Lengend Snippet: FIGURE 2 – CCR7 location in cell lines. Flow cytometry analysis of cell surface (filled bars) and cytoplasmic (open bars) CCR7 staining in the 14 CRC cell lines and the lymphoma cell line Granta-519 as posi- tive control for cell surface CCR7 (a). (b) shows a representative example of membranous CCR7 staining in the lymphoma cell line Granta-519 and the lack thereof in the CRC cell lines Colo205 and Colo320.

Article Snippet: CRC cell lines (CaCo2, Colo205, Colo206F, Colo320, Cx2, Cx94, DLD1, HCT116, HT29, LS174T, SW403, SW480, SW620, SW948, SW1116) and lymphoma cell lines (Granta519, Jeko1, KMH-2, Molt-4, Raji, SKW-3, SU-DHL4, U698, WSU-NL, YT) have been obtained from DSMZ (Germany, Braunschweig) and CLS (Germany, Eppelheim) and were cultured in RPMI 1640 medium (Biochrom, Berlin, Germany) supplemented with 10% FCS, glutamine and antibiotics.

Techniques: Flow Cytometry, Staining, Control

Journal: International journal of cancer

Article Title: Identification of truncated chemokine receptor 7 in human colorectal cancer unable to localize to the cell surface and unreactive to external ligands.

doi: 10.1002/ijc.23704

Figure Lengend Snippet: FIGURE 3 – Response to ligands CCL19/21. CCR7 functionality was analyzed by actin polymerization (a) and ERK-1/2 phosphoryla- tion (b, c) in the CRC cell lines Colo205 and Colo320, and the lym- phoma cell line Granta-519. (a) The F-actin content was determined by flow cytometry after stimulation with the CCR7-ligands CCL19, CCL21, and PMA as positive control. Results are expressed relative to the mean fluorescence intensity of the unstimulated samples. One rep- resentative out of 2 experiments is shown. (b) The ERK-1/2 activation was determined by antibodies recognizing the phosphorylated ERK-1/ 2 after incubation with 500 ng/ml CCL19/21 or PMA. Mean values and standard errors of 3 independent experiments are shown for optimized time points for each cell line. (c) Shows 1 representative experiment upon CCL19 incubation for Granta-519 and Colo320.

Article Snippet: CRC cell lines (CaCo2, Colo205, Colo206F, Colo320, Cx2, Cx94, DLD1, HCT116, HT29, LS174T, SW403, SW480, SW620, SW948, SW1116) and lymphoma cell lines (Granta519, Jeko1, KMH-2, Molt-4, Raji, SKW-3, SU-DHL4, U698, WSU-NL, YT) have been obtained from DSMZ (Germany, Braunschweig) and CLS (Germany, Eppelheim) and were cultured in RPMI 1640 medium (Biochrom, Berlin, Germany) supplemented with 10% FCS, glutamine and antibiotics.

Techniques: Cytometry, Positive Control, Activation Assay, Incubation

Journal: International journal of cancer

Article Title: Identification of truncated chemokine receptor 7 in human colorectal cancer unable to localize to the cell surface and unreactive to external ligands.

doi: 10.1002/ijc.23704

Figure Lengend Snippet: FIGURE 4 – Truncated CCR7 transcript in CRC cell lines. (a) dis- plays the CCR7 transcript (NM_001838). The signal peptide encoding sequence is displayed by the nucleotides (nt) 64-135 (grey bar). Sequencing analyses of CRC cell lines resulted in either a nonsense PCR product, a product with not defineable initial nucleotides of the signal peptide product (b) or an entire encoding product. Primers are shown as black arrows. Vertical lines display the exon transitions.

Article Snippet: CRC cell lines (CaCo2, Colo205, Colo206F, Colo320, Cx2, Cx94, DLD1, HCT116, HT29, LS174T, SW403, SW480, SW620, SW948, SW1116) and lymphoma cell lines (Granta519, Jeko1, KMH-2, Molt-4, Raji, SKW-3, SU-DHL4, U698, WSU-NL, YT) have been obtained from DSMZ (Germany, Braunschweig) and CLS (Germany, Eppelheim) and were cultured in RPMI 1640 medium (Biochrom, Berlin, Germany) supplemented with 10% FCS, glutamine and antibiotics.

Techniques: Sequencing

Journal: International journal of cancer

Article Title: Identification of truncated chemokine receptor 7 in human colorectal cancer unable to localize to the cell surface and unreactive to external ligands.

doi: 10.1002/ijc.23704

Figure Lengend Snippet: FIGURE 5 – RT-PCR Analyses of exon1/exon2 mRNA and subsequent mRNA level of CCR7. Ratios of CCR7 mRNA/ PBDG mRNA are displayed by box plots (25th, 50th and 75th percentile values) on the Y-axis in logarhythmic scale. Fragments (f0-f4) amplified by dif- ferent primers pairs are listed on the X- axis. (a) Shows three groups of CRC cell lines divided by their different pattern of CCR7 fragment levels. Group I with no/ marginal expression of fragment 0 (f0) and fragment 1 (f1), group II expressing up to 1log lower levels of f0 and f1 and group III displaying equivalent levels for all 4 fragments. (b) Shows the equivalent groups for CRC tissue samples. Group I with marginal/ up to 2log lower levels for fragment 0, 1 and 2, group II expressing up to 1log lower of fragments 0–2 and group III displaying equivalent levels for all 4 fragments. (c) As control experi- ments, CRC cell line- and CRC tissue- DNA were analyzed by using primers spanning the exon 1 and intron 1 bound- ary in comparison to a primer pair bind- ing exon3 (fragment 3). Furthermore, lymphoma cell lines (cDNA) were eval- uated for all the fragments investigated in the CRC cell lines (CRC cell lines-DNA (I), CRC tissues-DNA (II), lymphoma cell lines-cDNA (III).

Article Snippet: CRC cell lines (CaCo2, Colo205, Colo206F, Colo320, Cx2, Cx94, DLD1, HCT116, HT29, LS174T, SW403, SW480, SW620, SW948, SW1116) and lymphoma cell lines (Granta519, Jeko1, KMH-2, Molt-4, Raji, SKW-3, SU-DHL4, U698, WSU-NL, YT) have been obtained from DSMZ (Germany, Braunschweig) and CLS (Germany, Eppelheim) and were cultured in RPMI 1640 medium (Biochrom, Berlin, Germany) supplemented with 10% FCS, glutamine and antibiotics.

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Control, Comparison