Journal: Scientific Reports

Article Title: Impact of Escherichia coli K12 and O18:K1 on human platelets: Differential effects on platelet activation, RNAs and proteins

doi: 10.1038/s41598-018-34473-w

Figure Lengend Snippet: Western blot and ELISA analysis of HMBS and ATP2C1. ( a ) Cell lysates were applied to SDS-PAGE gels under reducing conditions. HMBS and ATP2C1 protein were detected in platelets using a polyclonal antibody. Incubation of platelets with E. coli K12 and E. coli O18:K1 in 1:5 or 1:10 platelet-bacteria ratios converted HMBS 47 kDa form to a 40 kDa protein. ATP2C1 was not affected. GAPDH was used as a loading control. ( b ) The releasates of the platelet-bacteria mix were collected after centrifugation (500 g, 10 minutes without break). HMBS, ATP2C1 and LRCH4 levels were measured by ELISA. Data represents the mean of three independent experiments (n = 3). ATP2C1 was detectable in platelet supernatants, while HMBS and LRCH4 proteins were either not released from platelets or in concentrations below the detection level of the ELISA (data not shown). BDL, below detection limit. Western blot results are representative image of three replications. The same exposure was applied equally across the entire image. The original pictures of the full-length western blots can be found in Supplementary Fig. .

Article Snippet: The blots were incubated overnight with primary anti-human antibodies (rabbit polyclonal ATP2C1 (1:750), LRCH4 (1:500), HMBS (1:1000) or GAPDH (1:600) from Atlas Antibodies, Bromma, Sweden).

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay, SDS Page, Incubation, Bacteria, Control, Centrifugation

Journal: Oncology Reports

Article Title: Long-term effects of short hairpin RNA-targeted human telomerase reverse transcriptase on suppression of SGC-7901 cell proliferation by inhibition of telomerase activity

doi: 10.3892/or.19.2.575

Figure Lengend Snippet: Figure 2. Effect of shRNA-targeted hTERT on hTERT gene expression. (A) Effect of shRNA-targeted hTERT on the level of hTERT mRNA expression. This was represented as NhTERT, which is the ratio of hTERT and GAPDH mRNA copies in each group. The different bands represent the mean of the results of the different groups. There was a significant difference between psi-hTERT1 or psi-hTERT2 and the psi-control; n=3 and *P<0.05 vs. psi-control. (B) Western blotting results demonstrated the presence of hTERT and GAPDH protein among the different groups. To standardize the protein quantity, GAPDH protein was also detected by Western blotting. The upper band is the hTERT protein, while the lower band is the GAPDH protein. (C) Effect of shRNA-targeted hTERT on the level of hTERT protein expression. The bands on the film were scanned by Typhoon 9400 phosphoimager and digitized using ImageQuant software. The ratio of the hTERT versus the GAPDH expression was used as a measurement of the hTERT level. The different bands represent the mean of the results of the different groups. There was a significant difference between psi-hTERT1 or psi-hTERT2 and the psi-control; n=3 and *P<0.05 vs. psi-control.

Article Snippet: The antibodies against hTERT and GAPDH were obtained from Boster Biological Technology (Wuhan, China) and horseradish peroxidase (HRP)-conjugated secondary antibody, from Jackson Immuno-Research Laboratories (West Grove, PA) and the electrochemiluminescence (ECL) reagents, from Amersham Biosciences (Buckinghamshire, UK).

Techniques: shRNA, Gene Expression, Expressing, Control, Western Blot, Software