Question: Based On These Results Provide 250 Words Abstract About It!

WASP and Verprolins WH1 BR GBD PRO V V C A N-WASP (human) WHI BR PRO V с Las17p (yeast) Abbreviations: WH1: GBD: PRO: V: WASP homology domain1; GTPase binding domain; Poly-proline; Verprolin-homology domain, Cofilin-homology or central domain; Acidic region; A: H440 a.a. WIRE (human) 817 a.a. Vrp1 (yeast) 4 Human N-WASP-WIRE Complex Cdc42 ACT1 ACT1 ARP2/3 N-WASP WHI BR GBD PRO V WIRE HH ACT1 Yeast Las 17-Vrp1 Complex ACT1 ARP2/3 Las 17 WHI BR PRO V Vrpl HH ACT1 N-WASP full length WHI CRIB Proline rich V V.CA N-WASP AVCA N-WASP AV V2 N-WASP AV N-WASP AWHI Fig. 2.2. Schematic of N-WASP deletion mutants used in this study WIRE full length V WASP binding do WIRE AV WIRE AC100 Fig. 2.3. Schematic of WIRE deletion mutants used in this study Viability of las 174 cells expressing N-VASP, WIRE and N-WASP+VIRE at 37°C WIRE AC100 Fig. 2.3. Schematic of WIRE deletion mutants used in this study Viability of las 174 cells expressing N-WASP, WIRE and N-WASP+WIRE at 37°C Wild Type Yeast can grow at 24°C and 37°C las 17A strain can grow at 24°C BUT not at 37°C Plasmids expressing N-WASP or WIRE are transformed into yeast Transformants are tested for growth at different temperatures 24′ c 37′ c las/74 lax/74 N.WASP lax/7A WIRE las 17A N-WASP WIRE Fig. 1 Viability at 24°C and 37°C of las 174 cells that were untransformed, las 174 cells that were transformed with plasmids expressing N-WASP, N-WASP + WIRE and WIRE. Strains were streaked for single colonies on YPUAD agar, incubated at either 24°C and 37°C and imaged after 3 days. Localization of WIRE-EGFP in the presence and absence of N-WASP Localization of N-WASP-EGFP in the presence and absence of WIRE N-WASP EGFP N-WASP EGFP WIRE N-WASP WIRE EGFP WIRE EGFP Fig. 2 The green dots are referred to as cortical patches las 174 cells were transformed with plasmids coding for N-WASP-EGFP, WIRE-EGFP, N-WASP-EGFP+ WIRE and WIRE-EGFP+N-WASP. Transformed cells were grown to exponential phase at 24°C. The fluorescent tagged proteins were imaged in living cells using fluorescence microscopy. Upper panels: FITC-fluorescence optics. Lower panels: DIC optics. Co-Localization of N-WASP-EGFP and WIRE-CFP N-WASP EGFP WIRE CFP N-WASP EGFP WIRE CFP Fig. 3 las 174 cells were transformed with plasmids encoding for N-WASP-EGFP, WIRE-CFP and N-WASP- EGFP+ WIRE- CFP. Transformed cells were grown to exponential phase at 24°C. EGFP and CFP were imaged in live cells using fluorescence microscopy. Upper panels: FITC-fluorescence optics. Lower panels: CFP fluorescence optics. Western Blot to detect the expression of WIRE-EGFP and N-WASP-EGFP EGFP and CFP were imaged in live cells using fluorescence microscopy. Upper panels: FITC-fluorescence optics. Lower panels: CFP fluorescence optics. Western Blot to detect the expression of WIRE-EGFP and N-WASP-EGFP N-WASP-EGFP WIRE-EGFP N-WASP-EGFP+ WIRE-EO 75 a-GFP SO a-Hex Fig. 4. Total protein extracts from las 1 74 cells transformed with plasmids coding for N-WASP-EGFP, WIRE-EGFP, N-WASP-EGFP and WIRE-EGFP were analyzed by Western Blot using anti-GFP (a-GFP) and anti-hexokinase (a-Hex) serum Checking the viability of las 17A cells expressing N-WASPAWH1 + WIRE, N-WASP+WIRE and N-WASP+WIREAC100 24° c 37° C las175 las 174 las 17 A N-WASP + A WHI WIRE las 174 las174 NWASP | NWASP + + WIRE WIRE AC100 Fig. 5 Viability at 24°C and 37°C of las 174 cells that were untransformed, and las 174 cells that were transformed with plasmids coding for N-WASPAWH1 + WIRE, N-WASP + WIRE and N-WASP + WIREAC100. Transformed cells were streaked for single colonies on YPUAD agar, incubated at either 24°C and 37°C and imaged after 3 days. Comparison of N-WASP-EGFP and N-WASPAWH1-EGFP localization N-WASP EGFP N-WASP AWHI EGFP Fig. 6 las 174 cells that were transformed with plasmids coding for N-WASP-EGFP and N-WASPAWH1-EGFP. Transformed cells were grown to exponential phase at 24°C. EGFP was imaged in living cells using fluorescence microscopy. Upper panels: FITC-fluorescence optics. Lower panels: DIC optics. Comparison of WIRE-EGFP and WIREAC100-EGFP localization WIRE EGFP WIRE AC100 EGFP Fig. 6 las 174 cells that were transformed with plasmids coding for N-WASP-EGFP and N-WASPAWHI-EGFP. Transformed cells were grown to exponential phase at 24°C. EGFP was imaged in living cells using fluorescence microscopy. Upper panels: FITC-fluorescence optics. Lower panels: DIC optics. Comparison of WIRE-EGFP and WIREAC100-EGFP localization WIRE EGFP WIRE AC100 EGFP Fig. 7. las 174 cells that were transformed with plasmids coding for WIRE-EGFP and WIREAC100-EGFP. Transformed cells were grown to exponential phase at 24°C. EGFP was imaged in living cells using fluorescence microscopy. Upper panels: FITC-fluorescence optics. Lower panels: DIC optics Checking the viability of las 17A cells expressing N-WASP+WIRE N-WASPAV1 + WIRE, N-WASPAV1V2 + WIRE, N-WASPAVCA+WIRE 24° c 37° c las 174 NWASP – WIRE NWASP AV, WIRE las/74 NWASP AVCA WIRE las/74 NWASP AV V, WIRE Fig. 8. Viability at 24°C and 37°C of las 174 cells that were transformed with plasmids coding for N-WASP + WIRE, N-WASPAV + WIRE, N-WASP AV, V,+ WIRE and N-WASPAVCA + WIRE. Strains were streaked for single colonies on YPUAD agar, incubated at either 24°C and 37°C and imaged after 3 days. 250 word Abstract Introduction Results Conclusion