Journal: medRxiv

Article Title: Genetic Insight into Birt-Hogg-Dubé syndrome in Indian patients reveals novel mutations in FLCN

doi: 10.1101/2021.10.31.21264930

Figure Lengend Snippet: Legend: Light blue boxes represent exons and green boxes are exons with pathogenic mutations in FLCN . Most pathogenic mutations were found between exons 10-13. Circles indicate females, squares males. Family numbers are denoted in grey boxes with patients and asymptomatic members harboring pathogenic FLCN mutations. Patient phenotypes are also indicated in red, blue and green colors in individual symbols, and asymptomatic carriers are denoted with a pink dot.

Article Snippet: Taqman Copy number assays for exon 4 (Hs01200751_cn), exon 8 (Hs01889931_cn), exon 13 (Hs01203178_cn) of FLCN and RNase P (as reference) were performed in real-time PCR instrument.

Techniques:

Journal: medRxiv

Article Title: Genetic Insight into Birt-Hogg-Dubé syndrome in Indian patients reveals novel mutations in FLCN

doi: 10.1101/2021.10.31.21264930

Figure Lengend Snippet: Legend: Green: N-terminal/Longin of FLCN, Brown: C-terminal of FLCN. Structures of homology modelled monomers of FLCN protein – i) wild-type FLCN with amino-acid residues which were affected by mutations; ii) stop-gain mutant FLCN affecting chain termination after Glu211 (i.e., p . Gln212*) ; iii) indel (11-nucleotide) mutant FLCN affecting chain termination after Leu383 (i.e., p . Val384Phe*2) ; iv) indel (single-nucleotide) mutant FLCN, altering the frame from His429 to Thr429, and subsequently creating a stop-codon after Leu435 in the model (i.e., p . His429Thr*39) ; v) four-nucleotide duplicate mutant FLCN, altering the frame and terminating the chain after His442 in the model (i.e., p . Ala445Ser*11) .

Article Snippet: Taqman Copy number assays for exon 4 (Hs01200751_cn), exon 8 (Hs01889931_cn), exon 13 (Hs01203178_cn) of FLCN and RNase P (as reference) were performed in real-time PCR instrument.

Techniques: Mutagenesis

Journal: Experimental & molecular medicine

Article Title: Epithelial folliculin is involved in airway inflammation in workers exposed to toluene diisocyanate.

doi: 10.1038/emm.2017.180

Figure Lengend Snippet: Figure 3 TDI-HSA conjugate-induced FLCN and IL-8 production from A549 cells. A549 cells were treated with TDI-HSA or mock-HSA (negative control) for 24 h. (a) The intracellular expression of FLCN was evaluated by western blotting. The levels of FLCN (b) and IL-8 (c) released to the culture supernatants were evaluated by ELISA. (d) Correlation of FLCN and IL-8 levels in the culture supernatants. (e) Effect of TDI-HSA conjugate on A549 cell viability was assessed using a Cell Counting Kit 8. *Po0.05, **Po0.01 obtained by one- way ANOVA with Bonferroni’s post hoc test. FLCN, folliculin; HAS, human serum albumin; IL, interleukin; NS, not significant.

Article Snippet: Serum-specific IgG (sIgG) and serum-specific (sIgE) antibodies to TDI-HSA were detected using a homemade enzyme-linked immunosorbent assay (ELISA), as previously described.9,10 ELISA to measure serum levels of FLCN and interleukin-8 Commercial ELISA kits were used to measure the serum levels of FLCN (CUSABIO Biotech, Wuhan, Hubei Province, China) and interleukin-8 (IL-8) (Endogen, Woburn, MA, USA), as well as the levels in cell culture supernatants, following the manufacturer’s protocols.

Techniques: Negative Control, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Cell Counting

Journal: Experimental & molecular medicine

Article Title: Epithelial folliculin is involved in airway inflammation in workers exposed to toluene diisocyanate.

doi: 10.1038/emm.2017.180

Figure Lengend Snippet: Figure 4 Peripheral blood neutrophils-induced FLCN expression and release from A549 cells. A549 cells (2 × 105 cells) were co- cultured with peripheral blood neutrophils for 24 h. (a) Intracellular FLCN expression was evaluated by western blotting. (b) The levels of FLCN released to the culture supernatants were measured by ELISA. *Po0.05, **Po0.01, ***Po0.001 vs control, obtained by one-way ANOVA with Bonferroni’s post hoc test. FLCN, folliculin.

Article Snippet: Serum-specific IgG (sIgG) and serum-specific (sIgE) antibodies to TDI-HSA were detected using a homemade enzyme-linked immunosorbent assay (ELISA), as previously described.9,10 ELISA to measure serum levels of FLCN and interleukin-8 Commercial ELISA kits were used to measure the serum levels of FLCN (CUSABIO Biotech, Wuhan, Hubei Province, China) and interleukin-8 (IL-8) (Endogen, Woburn, MA, USA), as well as the levels in cell culture supernatants, following the manufacturer’s protocols.

Techniques: Expressing, Cell Culture, Western Blot, Enzyme-linked Immunosorbent Assay, Control